EASY-nLC Series Troubleshooting and

EASY-nLC Series Troubleshooting and

Thermo

EASY-nLC Series

Troubleshooting and

Maintenance Guide

(Software version 3.1)

60053-97228 Revision B January 2013

© 2013 Thermo Fisher Scientific Inc. All rights reserved.

EASY-nLC, AFC, IFC, nanoViper, and PepMap are trademarks and Acclaim, Dionex, LCQ Fleet, LXQ, LTQ, and Xcalibur are registered trademarks of Thermo Fisher Scientific Inc.

PEEK is a trademark of Victrex PLC. PEEKsil is a trademark of SGE International Pty Ltd Corp.

This product uses FreeRTOS from www.freertos.org

(available upon request). FreeRTOS is a registered trademark of Real Time Engineers Ltd.

The following are registered trademarks in the United States: Advion and RePlay are registered trademarks of

Advion BioSystems, Inc. CMOSens is a registered trademark of Sensirion AG Corp. Duran is a registered trademark of Schott AG. PhotoMOS is a registered trademark of Panasonic Electric Works, Co., Ltd. Teflon is a registered trademark of E.I. du Pont de Nemours and Company. VICI and Valco are registered trademarks of

Valco Instruments Co., Inc.

The following are registered trademarks in the United States and other countries:

Microsoft and Windows are registered trademarks of the Microsoft Corporation. SSH is a registered trademark of Tectia Corporation. Linux is a registered trademark of Linus Torvalds (individual).

All other trademarks are the property of Thermo Fisher Scientific and its subsidiaries.

Thermo Fisher Scientific Inc. provides this document to its customers with a product purchase to use in the product operation. This document is copyright protected and any reproduction of the whole or any part of this document is strictly prohibited, except with the written authorization of Thermo Fisher Scientific Inc.

The contents of this document are subject to change without notice. All technical information in this document is for reference purposes only. System configurations and specifications in this document supersede all previous information received by the purchaser.

Thermo Fisher Scientific Inc. makes no representations that this document is complete, accurate or errorfree and assumes no responsibility and will not be liable for any errors, omissions, damage or loss that might result from any use of this document, even if the information in the document is followed properly.

This document is not part of any sales contract between Thermo Fisher Scientific Inc. and a purchaser. This document shall in no way govern or modify any Terms and Conditions of Sale, which Terms and Conditions of

Sale shall govern all conflicting information between the two documents.

Release history: Revision A, March 2012; Revision B, January 2013. Revision B includes information about replacing a PLF pump with a PLU pump, information about upgrading the touch-screen software, and improved troubleshooting procedures.

Hardware versions: EASY-nLC II and EASY-nLC 1000 instruments

Software version: Touch-screen software version 3.1

For Research Use Only. Not for use in diagnostic procedures.

Regulatory Compliance

Thermo Fisher Scientific performs complete testing and evaluation of its products to ensure full compliance with applicable domestic and international regulations. When the instrument is delivered to you, it meets all pertinent electromagnetic compatibility (EMC) and safety standards as described in the Declaration of Conformity.

Changes that you make to the instrument might void compliance with one or more of these EMC and safety standards.

Changes to the instrument include replacing a part or adding components, options, or peripherals not specifically authorized and qualified by Thermo Fisher Scientific. To ensure continued compliance with EMC and safety standards, replacement parts and additional components, options, and peripherals must be ordered from Thermo Fisher Scientific or one of its authorized representatives.

FCC Compliance Statement

THIS DEVICE COMPLIES WITH PART 15 OF THE FCC RULES. OPERATION IS SUBJECT TO

THE FOLLOWING TWO CONDITIONS: (1) THIS DEVICE MAY NOT CAUSE HARMFUL

INTERFERENCE, AND (2) THIS DEVICE MUST ACCEPT ANY INTERFERENCE RECEIVED,

INCLUDING INTERFERENCE THAT MAY CAUSE UNDESIRED OPERATION.

CAUTION Read and understand the various precautionary notes, signs, and symbols contained inside this manual pertaining to the safe use and operation of this product before using the device.

Notice on Lifting and Handling of

Thermo Scientific Instruments

For your safety, and in compliance with international regulations, the physical handling of this Thermo Fisher

Scientific instrument requires a team effort to lift and/or move the instrument. This instrument is too heavy and/or bulky for one person alone to handle safely.

Notice on the Proper Use of

Thermo Scientific Instruments

In compliance with international regulations: This instrument must be used in the manner specified by Thermo Fisher

Scientific to ensure protections provided by the instrument are not impaired. Deviations from specified instructions on the proper use of the instrument include changes to the system and part replacement. Accordingly, order replacement parts from Thermo Fisher Scientific or one of its authorized representatives.

Notice on the Susceptibility to Electromagnetic Transmissions

Your instrument is designed to work in a controlled electromagnetic environment. Do not use radio frequency transmitters, such as mobile phones, in close proximity to the instrument.

For manufacturing location, see the label on the instrument.

Declaration of Conformity for the EASY-nLC II System

U.S. Safety and EMC (Electromagnetic Compliance) Standards

Safety

This instrument has been reviewed for compliance with standard ANSI/UL 3101-1, “Electrical Equipment for

Laboratory Use; Part 1: General Requirements,” 1st Edition.

EMC

This device complies with Part 15 of the FCC Rules. Operation is subject to the following two conditions:

1. This device may not cause harmful interference, and

2. This device must accept any interference received, including interference that may cause undesired operation.

WARNING Changes or modification to this unit not expressly approved by the party responsible for compliance could void the user’s authority to operate the equipment.

Note

This equipment has been tested and found to comply with the limits for a Class A digital device, pursuant to Part

15 of the FCC Rules. These limits are designed to provide reasonable protection against harmful interference when the equipment is operated in a commercial environment. This equipment generates, uses, and can radiate radio frequency energy and, if not installed and used in accordance with the instruction manual, may cause harmful interference to radio communications. Operation of this equipment in a residential area is likely to cause harmful interference in which case users will be required to correct the interference at their own expense.

You must use shielded cables with this unit to ensure compliance with the Class A FCC limits.

Canadian Safety and EMC (Electromagnetic Compliance) Standards

Safety

This instrument has been reviewed for compliance with standard CAN/CSA-C22.2 No. 61010-1, Second edition -

“Safety Requirements for Electrical Equipment for Measurement, Control, and Laboratory Use; Part 1: General

Requirements.”

EMC

This Class A digital apparatus meets all requirements of the Canadian Interference-Causing Equipment Regulations.

European Safety and EMC (Electromagnetic Compliance) Standards

Application of Council

Directive(s)

2006/95/EEC “Low Voltage”: Intertek Group plc

Standard(s) to which conformity is declared

89/336/EEC “Electromagnetic Compatibility”: DELTA Denmark

EN61010-1:2001, Second edition - “Safety Requirements for Electrical Equipment for

Measurement, Control and Laboratory Use”

EN61010-2-81 “Part 2-081: Particular requirements for automatic and semi-automatic laboratory equipment for analysis and other purposes”

EN/(IEC) 61326-1:2006, A1(1998), A2(2001) and A3(2003) “EMC requirements for electrical equipment for measurement, control and laboratory use”

Manufacturer’s Name Proxeon Biosystems A/S

Manufacturer’s Address Edisonsvej 4, DK-5000 Odense, Denmark

Type of Equipment

Model Name

Model Numbers

Serial Number

Year of Manufacture

Laboratory Instrumentation

EASY-nLC™ II

LC110/LC111

LC-000100 and later

2010–

I, the undersigned hereby declare that the equipment specified above conforms to the above Directive(s) and

Standard(s).

Ole Vorm, Site Manager August 31, 2010

Declaration of Conformity for the EASY-nLC 1000 System

U.S. Safety and EMC (Electromagnetic Compliance) Standards

Safety

This instrument has been reviewed for compliance with standard UL 61010-1, “Electrical Equipment for

Measurement, Control, and Laboratory Use; Part 1: General Requirements.”

EMC

This device complies with Part 15 of the FCC Rules. Operation is subject to the following two conditions:

1. This device may not cause harmful interference, and

2. This device must accept any interference received, including interference that may cause undesired operation.

WARNING Changes or modification to this unit not expressly approved by the party responsible for compliance could void the user’s authority to operate the equipment.

Note

This equipment has been tested and found to comply with the limits for a Class A digital device, pursuant to Part

15 of the FCC Rules. These limits are designed to provide reasonable protection against harmful interference when the equipment is operated in a commercial environment. This equipment generates, uses, and can radiate radio frequency energy and, if not installed and used in accordance with the instruction manual, may cause harmful interference to radio communications. Operation of this equipment in a residential area is likely to cause harmful interference in which case users will be required to correct the interference at their own expense.

You must use shielded cables with this unit to ensure compliance with the Class A FCC limits.

Canadian Safety and EMC (Electromagnetic Compliance) Standards

Safety

This instrument has been reviewed for compliance with standard CAN/CSA-C22.2 No. 61010-1, Second edition -

“Safety Requirements for Electrical Equipment for Measurement, Control, and Laboratory Use; Part 1: General

Requirements.”

EMC

This Class A digital apparatus meets all requirements of the Canadian Interference-Causing Equipment Regulations.

European Safety and EMC (Electromagnetic Compliance) Standards

Application of Council

Directive(s)

Standard(s) to which conformity is declared

2004/108/EC “Electromagnetic Compatibility”

2006/95/EC “Low Voltage Directive (LVD)”

IEC 61010-1: Second edition, 2001. Safety Requirements for Electrical Equipment for

Measurement, Control and Laboratory Use. Also conformity with

• EN 61010-1:2001

• UL 61010-1, second edition

• CAN/CSA-C22.2 No. 61010-1, Second Edition

EN/(IEC) 61326-1:2006. Electrical equipment for measurement, control and laboratory use - EMC requirements - Part 1: General requirements

EN/(IEC) 61000-3-2:2006+A1+A2. Electromagnetic compatibility (EMC) - Part 3-2:

Limits - Limits for harmonic current emissions (equipment input current 16 A per phase)

EN/(IEC) 61000-3-3:2008. Electromagnetic compatibility (EMC) - Part 3-3: Limits -

Limitation of voltage changes, voltage fluctuations and flicker in public low-voltage supply systems, for equipment with rated current 16 A per phase and not subject to conditional connection

FCC CFR 47 Part 15, Class A. Radiated Emissions and Conducted Emissions

Manufacturer’s Name Proxeon Biosystems A/S

Manufacturer’s Address Edisonsvej 4, DK-5000 Odense, Denmark

Type of Equipment

Model Name

Model Numbers

Serial Number

Year of Manufacture

Laboratory Instrumentation

EASY-nLC™ 1000

LC120

LC-010000 and higher

2011–

I, the undersigned hereby declare that the equipment specified above conforms to the above Directive(s) and

Standard(s).

Ole Vorm, Site Manager July 19, 2011

WEEE Compliance

This product is required to comply with the European Union’s Waste Electrical & Electronic Equipment (WEEE)

Directive 2002/96/EC. It is marked with the following symbol:

Thermo Fisher Scientific has contracted with one or more recycling or disposal companies in each European Union

(EU) Member State, and these companies should dispose of or recycle this product. See www.thermoscientific.com/ rohsweee

 

for further information on Thermo Fisher Scientific’s compliance with these Directives and the recyclers in your country.

WEEE Konformität

Dieses Produkt muss die EU Waste Electrical & Electronic Equipment (WEEE) Richtlinie 2002/96/EC erfüllen.

Das Produkt ist durch folgendes Symbol gekennzeichnet:

Thermo Fisher Scientific hat Vereinbarungen mit Verwertungs-/Entsorgungsfirmen in allen EU-Mitgliedsstaaten getroffen, damit dieses Produkt durch diese Firmen wiederverwertet oder entsorgt werden kann. Mehr Information

über die Einhaltung dieser Anweisungen durch Thermo Fisher Scientific, über die Verwerter, und weitere Hinweise, die nützlich sind, um die Produkte zu identifizieren, die unter diese RoHS Anweisung fallen, finden sie unter www.thermoscientific.com/rohsweee .

Conformité DEEE

Ce produit doit être conforme à la directive européenne (2002/96/EC) des Déchets d'Equipements Electriques et

Electroniques (DEEE). Il est marqué par le symbole suivant:

Thermo Fisher Scientific s'est associé avec une ou plusieurs compagnies de recyclage dans chaque état membre de l’union européenne et ce produit devrait être collecté ou recyclé par celles-ci. Davantage d'informations sur la conformité de Thermo Fisher Scientific à ces directives, les recycleurs dans votre pays et les informations sur les produits Thermo Fisher Scientific qui peuvent aider la détection des substances sujettes à la directive RoHS sont disponibles sur www.thermoscientific.com/rohsweee.

CAUTION Symbol CAUTION VORSICHT ATTENTION PRECAUCION AVVERTENZA

Electric Shock: This instrument uses high voltages that can cause personal injury. Before servicing, shut down the instrument and disconnect the instrument from line power. Keep the top cover on while operating the instrument. Do not remove protective covers from PCBs.

Elektroschock: In diesem Gerät werden

Hochspannungen verwendet, die

Verletzungen verursachen können. Vor

Wartungsarbeiten muß das Gerät abgeschaltet und vom Netz getrennt werden. Betreiben Sie das Gerät nicht mit abgenommenem Deckel. Nehmen Sie die

Schutzabdeckung von Leiterplatten nicht ab.

Choc électrique: L’instrument utilise des tensions capables d’infliger des blessures corporelles. L’instrument doit être arrêté et débranché de la source de courant avant tout intervention. Ne pas utiliser l’instrument sans son couvercle. Ne pas enlever les étuis protecteurs des cartes de circuits imprimés.

Descarga eléctrica: Este instrumento utiliza altas tensiones, capaces de producir lesiones personales. Antes de dar servicio de mantenimiento al instrumento, éste debera apagarse y desconectarse de la línea de alimentacion eléctrica. No opere el instrumento sin sus cubiertas exteriores quitadas. No remueva las cubiertas protectoras de las tarjetas de circuito impreso.

Shock da folgorazione. L’apparecchio è alimentato da corrente ad alta tensione che puo provocare lesioni fisiche. Prima di effettuare qualsiasi intervento di manutenzione occorre spegnere ed isolare l’apparecchio dalla linea elettrica. Non attivare lo strumento senza lo schermo superiore. Non togliere i coperchi a protezione dalle schede di circuito stampato (PCB).

Chemical: This instrument might contain hazardous chemicals. Wear gloves when handling toxic, carcinogenic, mutagenic, or corrosive or irritant chemicals. Use approved containers and proper procedures to dispose waste oil.

Chemikalien: Dieses Gerät kann gefährliche Chemikalien enthalten. Tragen

Sie Schutzhandschuhe beim Umgang mit toxischen, karzinogenen, mutagenen oder

ätzenden/reizenden Chemikalien.

Entsorgen Sie verbrauchtes Öl entsprechend den Vorschriften in den vorgeschriebenen Behältern.

Chimique: Des produits chimiques dangereux peuvent se trouver dans l’instrument. Portez des gants pour manipuler tous produits chimiques toxiques, cancérigènes, mutagènes, ou corrosifs/irritants. Utiliser des récipients et des procédures homologuées pour se débarrasser des déchets d’huile.

Heat: Before servicing the instrument, allow any heated components to cool.

Hitze: Warten Sie erhitzte Komponenten erst nachdem diese sich abgekühlt haben.

Haute Temperature: Permettre aux composants chauffés de refroidir avant tout intervention.

Química: El instrumento puede contener productos quimicos peligrosos. Utilice guantes al manejar productos quimicos tóxicos, carcinogenos, mutagenos o corrosivos/irritantes. Utilice recipientes y procedimientos aprobados para deshacerse del aceite usado.

Prodotti chimici. Possibile presenza di sostanze chimiche pericolose nell’apparecchio. Indossare dei guanti per maneggiare prodotti chimici tossici, cancerogeni, mutageni, o corrosivi/irritanti. Utilizzare contenitori aprovo e seguire la procedura indicata per lo smaltimento dei residui di olio.

Altas temperaturas: Permita que los componentes se enfríen, ante de efectuar servicio de mantenimiento.

Calore. Attendere che i componenti riscaldati si raffreddino prima di effetturare l’intervento di manutenzione.

Fire: Use care when operating the system in the presence of flammable gases.

Feuer: Beachten Sie die einschlägigen

Vorsichtsmaßnahmen, wenn Sie das

System in Gegenwart von entzündbaren

Gasen betreiben.

Eye Hazard: Eye damage could occur from splattered chemicals or flying particles. Wear safety glasses when handling chemicals or servicing the instrument.

Verletzungsgefahr der Augen:

Verspritzte Chemikalien oder kleine

Partikel können Augenverletzungen verursachen. Tragen Sie beim Umgang mit

Chemikalien oder bei der Wartung des

Gerätes eine Schutzbrille.

Incendie: Agir avec précaution lors de l’utilisation du système en présence de gaz inflammables.

Fuego: Tenga cuidado al operar el sistema en presencia de gases inflamables.

Danger pour les yeux: Des projections chimiques, liquides, ou solides peuvent

être dangereuses pour les yeux. Porter des lunettes de protection lors de toute manipulation de produit chimique ou pour toute intervention sur l’instrument.

Peligro par los ojos: Las salicaduras de productos químicos o particulas que salten bruscamente pueden causar lesiones en los ojos. Utilice anteojos protectores al manipular productos químicos o al darle servicio de mantenimiento al instrumento.

Incendio. Adottare le dovute precauzioni quando si usa il sistema in presenza di gas infiammabili.

Pericolo per la vista. Gli schizzi di prodotti chimici o delle particelle presenti nell’aria potrebbero causare danni alla vista. Indossare occhiali protettivi quando si maneggiano prodotti chimici o si effettuano interventi di manutenzione sull’apparecchio.

General Hazard: A hazard is present that is not included in the above categories.

Also, this symbol appears on the instrument to refer the user to instructions in this manual.

When the safety of a procedure is questionable, contact your local Technical

Support organization for Thermo Fisher

Scientific San Jose Products.

Allgemeine Gefahr: Es besteht eine weitere Gefahr, die nicht in den vorstehenden Kategorien beschrieben ist.

Dieses Symbol wird im Handbuch außerdem dazu verwendet, um den

Benutzer auf Anweisungen hinzuweisen.

Wenn Sie sich über die Sicherheit eines

Verfahrens im unklaren sind, setzen Sie sich, bevor Sie fortfahren, mit Ihrer lokalen technischen

Unterstützungsorganisation für Thermo

Fisher Scientific San Jose Produkte in

Verbindung.

Danger général: Indique la présence d’un risque n’appartenant pas aux catégories citées plus haut. Ce symbole figure également sur l’instrument pour renvoyer l’utilisateur aux instructions du présent manuel.

Si la sûreté d’une procédure est incertaine, avant de continuer, contacter le plus proche Service Clientèle pour les produits de Thermo Fisher Scientific San

Jose.

Peligro general: Significa que existe un peligro no incluido en las categorias anteriores. Este simbolo también se utiliza en el instrumento par referir al usuario a las instrucciones contenidas en este manual.

Pericolo generico. Pericolo non compreso tra le precedenti categorie.

Questo simbolo è utilizzato inoltre sull’apparecchio per segnalare all’utente di consultare le istruzioni descritte nel presente manuale.

Cuando la certidumbre acerca de un procedimiento sea dudosa, antes de proseguir, pongase en contacto con la

Oficina de Asistencia Tecnica local para los productos de Thermo Fisher Scientific

San Jose.

Quando e in dubbio la misura di sicurezza per una procedura, prima di continuare, si prega di mettersi in contatto con il

Servizio di Assistenza Tecnica locale per i prodotti di Thermo Fisher Scientific San

Jose.

CAUTION Symbol CAUTION

Electric Shock: This instrument uses high voltages that can cause personal injury. Before servicing, shut down the instrument and disconnect the instrument from line power. Keep the top cover on while operating the instrument. Do not remove protective covers from PCBs.

Chemical: This instrument might contain hazardous chemicals. Wear gloves when handling toxic, carcinogenic, mutagenic, or corrosive or irritant chemicals. Use approved containers and proper procedures to dispose waste oil.

Heat: Before servicing the instrument, allow any heated components to cool.

Fire: Use care when operating the system in the presence of flammable gases.

Eye Hazard: Eye damage could occur from splattered chemicals or flying particles. Wear safety glasses when handling chemicals or servicing the instrument.

General Hazard: A hazard is present that is not included in the above categories.

Also, this symbol appears on the instrument to refer the user to instructions in this manual.

When the safety of a procedure is questionable, contact your local Technical

Support organization for Thermo Fisher

Scientific San Jose Products.

C

Contents

Preface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xix

Related Documentation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xx

Downloading Manuals from the Customer Manuals Web Site . . . . . . . . . . . . . xx

Safety and Special Notices . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .xxi

Contacting Us . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .xxiii

Chapter 1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .1

Hardware Components . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2

Autosampler . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2

Solvent System Components Behind the Right Side Panel . . . . . . . . . . . . . . . 5

Computer and Autosampler Behind the Left Side Panel . . . . . . . . . . . . . . . . 15

Back Panel. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16

Touch-Screen Application . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18

Description of the Touch-Screen Monitor. . . . . . . . . . . . . . . . . . . . . . . . . . . 18

Logging In to the EASY-nLC Instrument for Maintenance Tasks . . . . . . . . . 20

Closing Down the EASY-nLC Instrument . . . . . . . . . . . . . . . . . . . . . . . . . . 21

Specifications. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23

Performance Specifications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23

Technical Specifications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24

Chapter 2 Maintenance Schedule . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .27

Daily Maintenance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27

Weekly Maintenance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28

Quarterly Maintenance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28

Yearly Maintenance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29

Field Service Repairs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29

Thermo Scientific EASY-nLC Series Troubleshooting and Maintenance Guide

xiii

Contents

Chapter 3 Maintenance Scripts and Service Records . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .31

Maintenance Scripts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31

Prepare – Purge Solvent. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32

Prepare – Flush Air . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33

Prepare – Precolumn Equilibration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34

Prepare – Analytical Column Equilibration . . . . . . . . . . . . . . . . . . . . . . . . . . 36

Prepare – Isocratic Flow . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38

Test – MS Connection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39

Test – Sample Pickup . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42

Test – Leaks . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44

Test – Valve Check . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47

Test – Back Pressure . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 48

Test – Autosampler Torque. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49

Test – Pump Torque . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50

Calibrate – Valve Tune . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50

Calibrate – Flow Sensors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51

Calibrate – Reset Pressure Sensor. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 52

Calibrate – Direct Infusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 53

Keeping Service Records . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 55

Checking and Resetting the Device Usage Counters . . . . . . . . . . . . . . . . . . . . . 56

Chapter 4 Routine Maintenance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .57

Maintaining a Clean Working Environment . . . . . . . . . . . . . . . . . . . . . . . . . . . 58

Replacing the Main Power Fuse . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 58

Maintaining the Syringe Pumps . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60

Retracting the Piston . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 61

Replacing the Piston Seal and Cleaning the Piston in a PLF Pump . . . . . . . . 62

Replacing the Piston Seal in a PLU Pump . . . . . . . . . . . . . . . . . . . . . . . . . . . 67

Priming the Pump . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 71

Resetting the Pump Usage Counter. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72

Removing Air After Replacing a Piston Seal or a Pump . . . . . . . . . . . . . . . . . 73

Running the Leaks Script after Replacing a Piston Seal or a Pump . . . . . . . . 73

Maintaining the Rotary Valves . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 74

Cleaning the Rotor Seal and Stator . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 74

Replacing the Rotor Seal . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 77

Replacing the Stator . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 78

Replacing the Check Valves. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 80

Replacing an Inline Filter for the EASY-nLC 1000 Instrument . . . . . . . . . . . . . 81

Using nanoViper Fittings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 83

Replacing the Autosampler Needle . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 84

Replacing the Sample Loop . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 89

Replacing a Pressure Sensor for the PLU Pump . . . . . . . . . . . . . . . . . . . . . . . . . 90

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Thermo Scientific

Replacing a Flow Sensor . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 92

Replacing a Flow Sensor in the EASY-nLC 1000 Instrument . . . . . . . . . . . . 92

Replacing a Flow Sensor in the EASY-nLC II Instrument . . . . . . . . . . . . . . . 96

Replacing the Hard Drive . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 100

Managing the Devices List. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 105

Chapter 5 Field Service Maintenance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .107

Replacing the Built-In Computer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 108

Replacing the Monitor . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 113

Replacing a Pressure Sensor for a PLF Pump . . . . . . . . . . . . . . . . . . . . . . . . . . 117

Replacing the ASC Autosampler . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 122

Upgrading from an ASA Autosampler to an ASC Autosampler . . . . . . . . . . . . 128

Replacing the Autosampler Cooler . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 137

Replacing the ASC Cooler. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 137

Replacing the ASA Cooler . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 140

Replacing a Rotary Valve. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 147

Replacing a Pump . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 150

Replacing a PLU Pump . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 150

Replacing a PLF Pump with a PLU Pump. . . . . . . . . . . . . . . . . . . . . . . . . . 154

Replacing a Pump PCB . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 163

Replacing a Pump PCB in the PLU Pump . . . . . . . . . . . . . . . . . . . . . . . . . 163

Replacing a Pump PCB in the PLF Pump . . . . . . . . . . . . . . . . . . . . . . . . . . 167

Chapter 6 Troubleshooting. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .169

Troubleshooting Tips . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 170

Autosampler Problems. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 171

Contact Closure Problems . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 172

Instrument Startup Problems. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 172

Delayed Elution. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 173

Excessive Duration or Higher Pressure for the Column Equilibration and Sample Loading Steps . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 178

System Reaches Its Maximum Pressure During the Gradient. . . . . . . . . . . . 180

Sample Signal Weak or Absent . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 183

Slow or No Pressure Increase in Subsystem A or B . . . . . . . . . . . . . . . . . . . 187

Errors Reported by the Xcalibur Data System . . . . . . . . . . . . . . . . . . . . . . . 189

Carryover . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 194

Spray Issues . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 196

Chromatographic Performance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 197

Device Failures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 198

Miscellaneous . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 202

Using the Direct Controls for Troubleshooting and Maintenance . . . . . . . . . . 204

Using the Valve Controls. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 204

Using the Pump Controls . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 205

Using the Autosampler Controls . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 210

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Troubleshooting a Pump that Fails the Flush Air Script . . . . . . . . . . . . . . . . . 212

Troubleshooting a Pump that Fails the Leaks Script . . . . . . . . . . . . . . . . . . . . 216

Running a System Leak Test . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 218

Troubleshooting the Results of the System Leak Test . . . . . . . . . . . . . . . . . . . 221

Checking Valve S for a Leak . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 222

Checking the Sample Loop Connections for a Leak. . . . . . . . . . . . . . . . . . . 222

Locating a Leak in the Solvent Path from Valve B to Flow Sensor B . . . . . . 223

Locating a Leak in the Solvent Path from Pump A to Flow Sensor

A . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 224

Locating a Leak In or Between Valve S and Valve W. . . . . . . . . . . . . . . . . . 226

Locating a Leak Between the Flow Sensors and Valve S. . . . . . . . . . . . . . . . 234

Identifying a Leaking Check Valve . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 248

Troubleshooting a System Blockage . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 249

Running the Back Pressure Test for the A and B Solvent Paths . . . . . . . . . . 250

Troubleshooting a System Blockage when Test Solvent A Fails . . . . . . . . . . 251

Troubleshooting a System Blockage when Test Solvent B Fails . . . . . . . . . . 257

Troubleshooting the Autosampler Aspiration and Calibration. . . . . . . . . . . . . 261

Checking Sample Pickup. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 263

Troubleshooting Communication Problems . . . . . . . . . . . . . . . . . . . . . . . . . . 264

Network Access . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 264

Network Connection Failures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 265

Testing the Network Connection to the EASY-nLC Computer. . . . . . . . . . 266

Verifying that the LC/MS System Is Properly Grounded. . . . . . . . . . . . . . . . . 268

Chapter 7 Calibrating the Autosampler’s XYZ Robot . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .269

Replacing the Autosampler Adapter Plate . . . . . . . . . . . . . . . . . . . . . . . . . . . . 270

Replacing the Adapter Plate in the ASC Model Autosampler. . . . . . . . . . . . 271

Replacing the Adapter Plate in the ASA Model Autosampler . . . . . . . . . . . . 272

Managing Plate Formats . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 274

Selecting a Plate Format . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 274

Creating a New Plate Format . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 275

Deleting a Plate Format. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 277

Preparing the Autosampler for the Calibration Routines . . . . . . . . . . . . . . . . . 278

Calibrating Plates . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 280

Calibrating the Wash Bottle and Extra Vial Positions . . . . . . . . . . . . . . . . . . . 285

Chapter 8 Remote Support . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .287

Connecting the EASY-nLC Instrument to the Support Server. . . . . . . . . . . . . 288

Saving System Files on a USB Removable Storage Device . . . . . . . . . . . . . . . . 293

Downloading the Latest Firmware File . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 296

Chapter 9 Returning the EASY-nLC Instrument . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .299

Transport Instructions. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 299

Declaration of Contamination. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 301

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Appendix A Error Codes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .303

Appendix B Mobile Phase Viscosity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .311

Appendix C Consumables and Replacement Parts. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .313

Ordering Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 313

Solvent System Schematic for the EASY-nLC II Instrument . . . . . . . . . . . . . . 314

Solvent System Schematic for the EASY-nLC 1000 Instrument . . . . . . . . . . . 316

Common Replacement Parts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 318

Common Replacement Parts for the EASY-nLC II Instrument . . . . . . . . . . 318

Common Replacement Parts for the EASY-nLC 1000 Instrument . . . . . . . 319

Appendix D Quick Reference Guides for Routine Maintenance . . . . . . . . . . . . . . . . . . . . . . .321

Using nanoViper Fittings Quick Reference Guide. . . . . . . . . . . . . . . . . . . . . . 323

Connecting nanoViper Fittings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 323

nanoViper Tubing Connections . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 324

Returning the System to Atmospheric Pressure . . . . . . . . . . . . . . . . . . . . . . 325

Maintaining the PLU Pump Quick Reference Guide . . . . . . . . . . . . . . . . . . . 327

Maintaining the PLF Pump Quick Reference Guide . . . . . . . . . . . . . . . . . . . . 329

Maintaining the Rotary Valves Quick Reference Guide. . . . . . . . . . . . . . . . . . 331

Replacing a Damaged Autosampler Needle . . . . . . . . . . . . . . . . . . . . . . . . . . . 333

Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .335

Thermo Scientific EASY-nLC Series Troubleshooting and Maintenance Guide

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P

Preface

This guide describes the maintenance and troubleshooting procedures required to maintain the EASY-nLC™ instrument in optimal working condition.

The hardware maintenance procedures are divided into two chapters: Routine Maintenance and Advanced Maintenance. The Routine Maintenance chapter contains common maintenance procedures that you can perform to keep the EASY-nLC instrument in optimal working condition. The Advanced Maintenance chapter contains hardware repair procedures that can only be performed by a Thermo Fisher Scientific field service engineer or a skilled technician with equivalent training.

For information about setting up the EASY-nLC instrument and running chromatographic separations, refer to the EASY-nLC Series Getting Started Guide.

Contents

Related Documentation

Downloading Manuals from the Customer Manuals Web Site

Safety and Special Notices

Contacting Us

To suggest changes to documentation or to Help

If you are viewing a PDF file of this manual online, complete a brief survey about this document by clicking the button below.

If you are reading a printed copy of this manual, complete a brief survey about this document by going to the following URL.

http://www.surveymonkey.com/s/PQM6P62

Thermo Scientific EASY-nLC Series Troubleshooting and Maintenance Guide

xix

Preface

Related Documentation

In addition to this guide, Thermo Fisher Scientific provides the following documents for the

EASY-nLC instrument:

EASY-nLC Series User Guide for the Xcalibur 2.x Data System

EASY-nLC Preinstallation Requirements Guide

• EASY-nLC Series Getting Started Guide

Downloading Manuals from the Customer Manuals Web Site

For Thermo Scientific software and hardware products, Thermo Fisher Scientific provides instructional guides as PDF files on its Customer Manuals Web site.

Note If you are using a Thermo Scientific application to control the EASY-nLC system as part of an LC/MS system, the EASY-nLC manuals are also available from the data system computer’s Start menu.

To download the EASY-nLC manuals from the Customer Manuals Web site

1. Go to mssupport.thermo.com

.

2. On the Terms and Conditions page, click I Agree.

3. Click Customer Manuals in the left margin of the window.

4. On the right side of the window, click Search.

The Documents - Advanced Search page opens.

5. Open a list of linked PDF documents for the EASY-nLC as follows: a. In the Model list, select EASY-nLC and in the Document Type list, select Manual.

b. Click Submit Search.

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EASY-nLC Series Troubleshooting and Maintenance Guide Thermo Scientific

Preface

6. On the Documents page, click the link to the right of Download to view the PDF.

7. Click the Save a Copy icon, , browse to an appropriate folder, and then click Save to save a copy of the document.

Safety and Special Notices

Make sure to follow the safety practices presented in this guide, and observe the safety and special notices that appear in boxes.

Observe all written safety precautions during all phases of operation, service, and repair of this instrument. Failure to comply with these precautions or with specific warnings elsewhere in this manual violates safety standards of design, manufacture, and intended use of the instrument and might result in injury or loss of life.

The following two stickers appear on the EASY-nLC instrument:

This sticker warns you that the instrument includes a sharp needle and moving parts that are accessible to the operator. To prevent personal injury or damage to parts of the EASY-nLC instrument, take care when loading samples into the instrument’s tray compartment.

WARNING:

AVERTISSEMENT:

SHARP NEEDLE AND MOVING PARTS.

KEEP HANDS CLEAR.

AIGUILLE POINTURE ET PARTIES MOUVANTES.

` ´

This sticker alerts you to consult this manual for instructions on how to operate the instrument.

The safety and special notices in the documentation include the following:

CAUTION Highlights hazards to humans, property, or the environment. Each CAUTION notice is accompanied by an appropriate CAUTION symbol.

CAUTION Highlights electrical hazards to humans or property.

Thermo Scientific EASY-nLC Series Troubleshooting and Maintenance Guide

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Preface

CAUTION Highlights a sharp object hazard to humans.

CAUTION Highlights an eye hazard to humans.

CAUTION Highlights a chemical hazard to humans, property, or the environment.

CAUTION Highlights lifting hazards.

IMPORTANT Highlights information necessary to prevent damage to software, loss of data, or invalid test results; or might contain information that is critical for optimal performance of the system.

Note Highlights information of general interest.

Tip Highlights helpful information that can make a task easier.

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EASY-nLC Series Troubleshooting and Maintenance Guide Thermo Scientific

Preface

Contacting Us

There are several ways to contact Thermo Fisher Scientific for the information you need.

To contact Technical Support for the EASY-nLC instrument

Web site

E-mail

Address www.proxeon.com

(North and South America) [email protected]

(Other continents) [email protected]

Thermo Fisher Scientific

Edisonvej 4

DK-5000 Odense C

Find software updates and utilities to download at mssupport.thermo.com

.

To order consumable and spare parts for the EASY-nLC instrument

For the EASY-nLC 1000 instrument, go to: www.proxeon.com/productrange/nano_lc_easy-nlc_1000/accessories_spares/index.html

For the EASY-nLC II instrument, go to: www.proxeon.com/productrange/nano_lc/accessories-spares/index.html

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1

Introduction

The EASY-nLC instrument (see Figure 1 ) consists of a high-pressure dual pump

chromatography system that provides split-free flows down to the low nanoLiter/min range, an automated sampling system, and a built-in computer with a touch-screen interface for instrument control and troubleshooting. The automated sampling system includes a tray compartment that can hold standard sample vials and microtiter plates, an XYZ robot that moves to the sample location, and a syringe pump that draws only the user-specified sample volume.

The EASY-nLC II instrument operates in the high-performance pressure range of 1 to

300 bar and the EASY-nLC 1000 instrument operates in the ultra-high-performance pressure range of 1 to 1000 bar.

Figure 1.

EASY-nLC II instrument on the left and EASY-nLC 1000 instrument on the right

Thermo Scientific

To familiarize yourself with the EASY-nLC instrument and its instrument control software, review these topics.

Contents

Hardware Components

Touch-Screen Application

Specifications

Note For information about setting up the EASY-nLC instrument, installing the columns, creating methods, and running batches, refer to the EASY-nLC Series Getting

Started Guide.

EASY-nLC Series Troubleshooting and Maintenance Guide

1

1

Introduction

Hardware Components

Hardware Components

These topics describe the hardware components of the EASY-nLC instrument:

Autosampler

Solvent System Components Behind the Right Side Panel

Computer and Autosampler Behind the Left Side Panel

Back Panel

Autosampler

The autosampler module of the EASY-nLC instrument is located behind the tray compartment door. You can open the tray compartment door manually or by using the touch-screen controls.

With the use of a vial adapter plate or a microwell adapter plate, the tray compartment can hold sample vials, a 96-well microplate, or a 384-well microplate.

The robotic unit moves the z-axis needle holder along the x and y axes to the user-specified sample location during a run. When the autosampler needle reaches the sample location, it descends into the sample vial or microplate well.

IMPORTANT Because they can block the movement of the z-axis needle holder, avoid placing tall objects into the tray compartment.

The autosampler needle draws solvent from wash bottle W3 to perform a standard wash and ejects the solvent into wash bottle W4, which contains the needle wash insert. The autosampler can also perform a custom wash using the solvents from wash bottles W1–W3.

The waste beaker to the right of the tray compartment collects solvent that is delivered by pumps A and B when these pumps empty solvent to waste. This waste beaker also collects solvent delivered by pump A during the precolumn equilibration and sample loading steps

(for a two-column setup).

The EASY-nLC Series includes two supported versions of the autosampler module:

“ASC Model—Current” on page 3

“ASA Model—Legacy EASY-nLC II Instruments” on page 4

Note The ASC model has an integrated cooling compartment, which ensures stable temperature conditions throughout the entire vial plate. By contrast, the ASA model is cooled from underneath only.

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EASY-nLC Series Troubleshooting and Maintenance Guide Thermo Scientific

1

Introduction

Hardware Components

The following differences between the two autosampler models affect the routine calibration and maintenance procedures:

• The z-axis needle holders and the autosampler needles. This difference affects the autosampler needle replacement procedure.

• The location of the wash bottles. This difference affects the autosampler calibration procedure.

• The adapter plate holder. The adapter plate holder for the ASC model holds an additional six sample vials. This difference affects the autosampler calibration procedure and the maximum number of sample vials that you can run without reloading the autosampler.

ASC Model—Current

Figure 2

shows the ASC autosampler model that is installed in the current versions of the

EASY-nLC II and EASY-nLC 1000 instruments.

Figure 2.

ASC autosampler model installed in the current versions of the EASY-nLC instruments

y axis (left and right)

z-axis needle holder

Sample vial tray

x axis (in and out)

Thermo Scientific EASY-nLC Series Troubleshooting and Maintenance Guide

3

1

Introduction

Hardware Components

Figure 3

shows the tray compartment for the ASC autosampler as viewed from above when the tray compartment door is open. The tray compartment of the ASC autosampler model holds a vial adapter or a well plate adapter, six additional vials, three wash solvent bottles, and a wash bottle with a wash insert for cleaning the autosampler needle.

Figure 3.

Tray compartment for the ASC autosampler model (viewed from above)

Well plate (96 or 384) or vial (6 × 8) adapter

Three positions for wash solvents

Waste bottle with wash insert for cleaning of the outside of the needle

Six extra vial positions for standards or regular samples

ASA Model—Legacy EASY-nLC II Instruments

Figure 4

shows the ASA autosampler model that is installed in earlier versions of the

EASY-nLC II instrument.

Figure 4.

ASA autosampler model installed in earlier versions of the EASY-nLC II instrument

y axis (left and right)

z-axis needle holder

Adapter plate holder with peltier cooling

Wash bottles

x axis (in and out)

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EASY-nLC Series Troubleshooting and Maintenance Guide Thermo Scientific

1

Introduction

Hardware Components

Figure 5

shows the tray compartment for the ASA autosampler model as viewed from above when the tray compartment door is open. The tray compartment of the ASA autosampler model holds a vial adapter or a well plate adapter, three wash solvent bottles, and a wash bottle with a wash insert for cleaning the autosampler needle. The ASA autosampler model does not hold six additional vials.

Figure 5.

Tray compartment for the ASA autosampler model (viewed from above)

Solvent System Components Behind the Right Side Panel

The EASY-nLC solvent delivery system is located behind the right side panel and consists of three syringe pumps, three pressure sensors, two flow sensors, four rotary valves, four check valves (two for each assembly), and a mixing Tee.

Note Two of the syringe pumps (pumps A and B) produce the binary mobile phase and the third syringe pump (pump S) draws the sample into the sample loop that is attached to one of the rotary valves (valve S).

These topics describe the hardware components behind the right side panel:

View Behind the Right Side Panel

Syringe Pumps

Pressure Sensors

Flow Sensors

Check Valve Assemblies

Six-Port Rotary Valves

Autosampler Needle

View Behind the Right Side Panel

You can access the solvent system components by removing the instrument’s right side panel, which is secured to the housing with three captive quarter-turn screws.

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Hardware Components

Figure 6

shows the EASY-nLC II solvent system.

Figure 6.

Solvent system components behind the right panel of the EASY-nLC II instrument

A and B

Waste In line

Autosampler needle

Column Out line

Flow sensors

Mixing Tee

Valve W

2

1

6

4

Valve S

2

1

6

4

Valve A

2

1

6

4

Valve B

2

1

6

4

+

Pump S Pump A Pump B

Solvent A check valve assembly

Solvent B check valve assembly

Rotary valves

W, S, A, and B

Pressure sensors

S, A, and B

Pumps

S, A, and B

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EASY-nLC Series Troubleshooting and Maintenance Guide Thermo Scientific

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Introduction

Hardware Components

Figure 7

shows the solvent system components for the EASY-nLC 1000 instrument.

Figure 7.

Solvent system components behind the right panel of the EASY-nLC 1000 instrument

Solvent A check valve assembly

Solvent B check valve assembly

Autosampler needle

Four 3-position,

6-port valves

Pressure sensor S

Mixing Tee

Three syringe pumps

Pressure sensors

A and B

Flow sensors

A and B

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Introduction

Hardware Components

Syringe Pumps

The EASY-nLC instrument contains three syringe pumps. The binary liquid chromatography system consists of two high-pressure pumps: pumps A and B. The automated sampling system uses syringe pump S to draw sample from a sample vial or well and load it into the sample loop attached to valve S.

Figure 8

shows the EASY-nLC syringe pump components. The PLF model is installed in the

EASY-nLC II instrument. The PLU model is installed in the EASY-nLC 1000 instrument and is supplied as spare part for both instruments.

Each syringe pump consists of these primary components: a stepper motor, a pump body, a pump head, a piston, a piston seal, and a backup ring. The PLU model also has an LED panel and a pump printed circuit board (PCB) cover. The piston seal is the only user-replaceable component.

The pump head holds up to 140 μL of fluid. The backward movement of the piston draws solvent into the pump head and the forward movement of the piston pushes solvent into the attached high-pressure solvent line. The stepper motor controls the forward and backward movement of the piston within the pump head. The pump body provides piston alignment.

The piston seal allows the piston to move freely within the pump head. The piston seal is made of an extremely strong thermoplastic. The seal for the EASY-nLC 1000 pumps contains a wire spring that forces the inside flange of the seal against the piston to prevent leaks. The seals are not leak proof. In fact, wetting the surface of the pistons requires a small quantity of liquid, which acts as a lubricant to reduce wear on the piston seal. Through continued use and over time, the seal degrades and must be replaced. Allowing the pump to run dry and neglecting to rinse the pump head assemblies after pumping buffered eluents shorten the lifespan of the seals.

The backup ring holds the piston seal in place. It also provides better alignment of the piston.

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Introduction

Hardware Components

Figure 8.

Syringe pump components (showing the pump head removed from the pump body)

EASY-nLC II syringe pump (PLF model)

EASY-nLC 1000 syringe pump (PLU model)

Thermo Scientific

Pump head

Piston seal

Piston

Backup ring

Rotary switch

Pump body

Stepper motor

LED panel

Backup ring

Motor cable

The syringe pumps perform these functions.

Component

Pumps A and B

Pump S

Function

Draw solvent from the solvent bottles on top of the instrument, empty solvent to the waste beaker, and deliver solvent to the mixing Tee.

Draws solvent from the W3 bottle, draws sample from the specified sample location into the autosampler needle, and empties solvent to the W4 bottle in the autosampler tray compartment.

For maintenance purposes, the system tracks the volume pumped by each pump

(see

“Checking and Resetting the Device Usage Counters” on page 56

).

The PLF model pumps installed in the EASY-nLC instrument have an error LED. The LED illuminates green when the instrument is turned on and the pump is functioning properly.

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Hardware Components

The LED panels for the PLU model pumps installed in the EASY-nLC 1000 instrument provide the following status information.

LED

Flow sensor for

Pumps A and B

Power

Error

States

Illuminates blue—the instrument is turned on and the flow sensor is connected to the pump.

Not illuminated—the instrument is not turned on or the flow sensor is not connected to the pump.

Illuminates yellow—The instrument power is turned on.

Not illuminated—The instrument power is turned off.

Illuminates red—A pump error has occurred.

Not illuminated—The pump is operating properly.

Pressure Sensors

For pumps A, B, and S, the three pressure sensors connect the solvent path between the pump outlet and the associated rotary valve.

Using the strain gauge principle to measure pressure accurately, the pressure sensors are flow-through, zero dead volume devices with a wetted path that contains 12 μL of swept volume. The pressure sensors for the EASY-nLC II instrument have an operating range of 0 to 5000 psi (345 bar), and the pressure sensors for the EASY-nLC 1000 instrument have an operating range of 0 to 15 000 psi (1034 bar).

Flow Sensors

The two flow sensors provide feedback control for pumps A and B. The flow sensors are liquid mass flow meters with a CMOSens™ patented technology. Each flow meter consists of a length of fused silica capillary, a sensor chip, a heating resistor, and two temperature sensors

(see

Figure 9

).

Figure 9.

Flow sensor schematic

Flow

Tubing

Sensor chip

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EASY-nLC Series Troubleshooting and Maintenance Guide

Heater

Temperature sensors

Based on CMOSens™ Technology

Principle patented: US Pat. 6 813 944

Thermo Scientific

1

Introduction

Hardware Components

The flow sensors determine the flow rate of solvents passing through the system by measuring the heat transfer through the fused silica capillary. Outside the capillary, a heating resistor on a thermally optimized membrane is maintained above ambient temperature. When liquid flows through the capillary, the temperature distribution upstream and downstream of the heating resistor is disturbed. The two temperature sensors measure this temperature asymmetry.

Because the temperature distribution depends on both the solvent flow rate and the solvent mixture, you must recalibrate the flow sensors whenever you change the solvent types for solvents A and B. For example, if you use methanol instead of acetonitrile for the solvent B

mixture, run the script “Calibrate – Flow Sensors” on page 51 .

The EASY-nLC 1000 instrument uses two flow sensors with a maximum measuring range of 5.0 μL/min. The EASY-nLC II instrument uses two different flow sensors. The maximum measuring range is approximately 2.9 μL/min for flow sensor A and 4.5 μL/min for flow sensor B.

The wetted path for the EASY-nLC 1000 instrument includes external inline filters for the flow sensor inlets and outlets. When the inline filters become clogged, they must be replaced.

For the EASY-nLC II instrument, the tubing that connects the valve to the flow sensor contains a built-in filter.

Check Valve Assemblies

When the corresponding high-pressure (6-port rotary) valves for pumps A or B are in position 1–2, the check valves perform these functions:

• The solvent-side check valve prevents backflow into the associated solvent inlet line when the associated pump is emptying solvent to the waste beaker.

• The waste-side check valve prevents backflow from the waste beaker when the associated pump is filling and drawing solvent from the associated solvent bottle.

Figure 10. Check valve connections

From the solvent reservoir bottle on the top of the instrument

Solvent-side check valve

Connected to port 2 of valve A or B

Waste-side check valve

Thermo Scientific

Routed to the waste beaker

The check valves are replaceable. For more information about determining if the check valves are leaking, see

“Identifying a Leaking Check Valve” on page 248

.

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Hardware Components

Six-Port Rotary Valves

The three-position, six-port rotary valves consist of a driver, a rotor seal, and a stator (see

Figure 11 ).

Figure 11. Valve components

Driver

Rotor seal

Stator

Screws that secure the stator to the driver

The six-port valves can be in three different positions: 1–2 position, 1–6 position, and centered (see

Figure 12

).

Figure 12. Valve positions

1–2 position 1–6 position Centered

2

3

1

6

4 5

2

3

1

6

4 5

For maintenance purposes, the system tracks the number of valve shifts for each valve

(see

“Checking and Resetting the Device Usage Counters” on page 56

).

Depending on the valve position, the valves perform the functions listed in

Table 1 .

Note The EASY-nLC solvent system contains three subsystems, one for each pump.

A subsystem includes the pump, the pressure sensor, the solvent lines to and from the pressure sensor, and the rotary valve.

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Thermo Scientific

1

Introduction

Hardware Components

.

Table 1. Valve functions

Component Function

Valves A and B Direct the solvent flow from syringe pump A and B, respectively.

1–2 position Directs the solvent flow toward the check valve. When the valve is in the 1–2 position, the pump draws solvent from the solvent bottle on top of the instrument or empties solvent to the waste beaker.

1–6 position Directs the solvent flow toward the mixing Tee.

Centered • Prevents backflow into the solvent line upstream of the mixing Tee.

Valve S

• Allows the back pressure for the solvent A and B lines to equalize before the gradient step of a sample run.

• Blocks the solvent path through the valve and to the subsystem for several maintenance scripts.

Directs the solvent flow from syringe pump S and the solvent flow from the mixing Tee.

1–2 position The solvent flow bypasses the sample loop.

1–6 position The solvent flow passes through the sample loop.

Centered • Isolates the pump S subsystem for the Flush Air script.

• Blocks the solvent path after valve S for the system leak test (Leaks script with the System check box selected).

Valve W

• Shuts off the flow to the Column Out line and provides back pressure during the Flow Sensor

Calibration script.

Directs the solvent flow through the venting Tee to the waste beaker or provides pressure venting to the system.

1–2 position Directs the solvent flow toward the column by blocking the solvent path through valve W.

1–6 position Directs the solvent flow from the venting Tee to the waste beaker.

Centered Blocks the solvent flow through the valve to the waste beaker.

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Hardware Components

Autosampler Needle

During a sample run, the autosampler needle descends into the requested sample vial or well, and then pump S draws the requested sample volume into the needle tubing. The autosampler needle is a user-replaceable part. For information about replacing the

autosampler needle, see “Replacing the Autosampler Needle” on page 84 .

The autosampler needle for the ASC autosampler is a length of PEEKsil™ tubing with a sleeve and a stainless steel fitting at one end and a PEEK™ nut and plastic stop at the other end

(see

Figure 13 ). Fitting into a slot in the panel between the solvent system compartment and

the tray compartment, the needle retainer properly positions the needle tubing.

Figure 13. Autosampler needle for the ASC autosampler (current EASY-nLC II and EASY-nLC 1000 instruments)

Stainless steel fitting and sleeve

PEEK nut

Needle retainer

Plastic stop

Figure 14 shows autosampler needle for the ASA autosampler.

Figure 14. Autosampler needle for the ASA autosampler model (earlier EASY-nLC II instruments)

For both needles, the stainless steel fitting connects to port 1 of valve S. The tubing routes through a cutout in the panel that separates the solvent delivery system components from the autosampler compartment. The other end of the tubing slides into the z-axis holder and is secured with a PEEK nut.

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Introduction

Hardware Components

Computer and Autosampler Behind the Left Side Panel

You can access the integrated computer and the autosampler needle fitting that connects to the needle guide by removing the left side panel of the instrument (see

Figure 15 ). Three

captive quarter-turn screws secure the left side panel to the instrument housing.

Figure 15. Internal features (behind the left side panel)

Integrated computer

Needle tubing

Needle guide

Thermo Scientific

Autosampler

Enclosed refrigerated sample compartment

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Hardware Components

Back Panel

The system communication, contact closure, and power line connections are on the back panel of the instrument (see

Figure 16 and Figure 17

).

Figure 16. Back panel of the EASY-nLC instrument (prior to March 2012)

PRO EON

P-BUS

IN3 OUT3 IN2 OUT2 IN1 OUT1

PS/2 LAN

HARD

DRIVE

This device complies with Part 15 of the FCC Rules.

Operation is subject to the following two conditions:

(1) This device may not cause harmful interference, and

(2) This device must accept any interference received, including

interference that may cause undersired operation.

This instrument is for research use only.

DEVICE: EASY-NLC

P/N LC100

LAN MAC ADDRESS: 00-6--E0-45-4E-28

Proxeon Biosystems A/S

Web: www.proxeon.com

Phone +45 6557 2300 Fax +45 6557 2301

MONITOR

I

O

110-120V▼

220-240▼

120/230 V : 50/60 Hz: 250 W

Fuse at 120 V– : T5 AL 250 V

Fuse at 230 V– : T2.5 AL 250 V

Figure 17. Back panel of the EASY-nLC instrument (with the latest computer box)

P-BUS

IN3 OUT3 IN2 OUT2 IN1 OUT1

USB LAN RS-232

HARD

DRIVE

This device complies with Part 15 of the FCC Rules.

Operation is subject to the following two conditions:

(1) This device may not cause harmful interference, and

(2) This device must accept any interference received, including

interference that may cause undersired operation.

This instrument is for research use only.

DEVICE: EASY-NLC

P/N LC100

LAN MAC ADDRESS: 00-6--E0-45-4E-28

Thermo Fisher Scientific A/S

Web: www.proxeon.com

Phone +45 6557 2300 Fax +45 6557 2301

MONITOR

I

O

110-120V▼

220-240▼

120/230 V : 50/60 Hz: 250 W

Fuse at 120 V– : T5 AL 250 V

Fuse at 230 V– : T2.5 AL 250 V

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Hardware Components

Table 2 lists the function of each back panel element.

Table 2. Back panel elements

Element

RS-232

USB

Description

For communication with add-on devices, for example a syringe drive

Port for keyboard or mouse or removable USB storage device, two (2) provided

Input for connection of keyboard and mouse PS/2 (discontinued computer box only)

LAN For 10/100 MB/sec Ethernet connection

MONITOR Output for connection of external display

P-BUS

IN/OUT pins

For communication with add-on devices

Contact closure (primarily for communication with the mass spectrometer)

Power switch I|O

Fuse holder below

I|O switch

P/N

S/N

Replaceable fuse ratings:

• For 120 V, T 5 AL, 250 V

• For 230 V, T 2.5 AL, 250 V

Product order code

Instrument serial number

LAN MAC ADDRESS MAC address of the embedded computer (Gives the EASY-nLC computer a unique network identifier on your local network.)

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Touch-Screen Application

Touch-Screen Application

The EASY-nLC instrument includes an integrated computer with instrument control software that you access from the instrument monitor.

These topics provide an overview of the touch-screen application and describe how to log in to the EASY-nLC instrument as an administrator and how to close down the instrument:

Description of the Touch-Screen Monitor

Logging In to the EASY-nLC Instrument for Maintenance Tasks

Closing Down the EASY-nLC Instrument

Description of the Touch-Screen Monitor

The instrument monitor is a transparent layer of glass that detects finger pressure (even while wearing gloves) and sends the corresponding commands to the computer (see

Figure 18 ).

Figure 18. Touch-screen controls

Home > Overview page

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Introduction

Touch-Screen Application

Instrument control from this touch-screen application includes a method wizard that helps you create chromatographic methods, a batch editor that helps you create and run sample batches (injection sequences); maintenance scripts and record keeping features; and direct control of the valves, pumps, XYZ robot, and tray temperature.

Note This guide describes how to log in to the instrument as an administrator, how to use the maintenance scripts and record keeping features, and how to exit the touch-screen application in a controlled manner and turn off the instrument before performing maintenance procedures on the instrument’s hardware components.

For information about the maintenance scripts and record keeping features, see

Chapter 3, “Maintenance Scripts and Service Records.” For information about creating

methods and sample batches and running sample batches, refer to the EASY-nLC Series

Getting Started Guide (for the touch-screen application).

The EASY-nLC touch-screen application contains five menu tabs at the top of the touch screen. When you press a tab at the top of the touch screen, a set of side tabs appears

on the left side of the touch screen. Figure 19 shows the menu layout for the Home >

Overview page.

Figure 19. EASY-nLC touch screen with the menu layout on the Home > Overview page

Top menu tabs

Logo icon

Side tabs

Current user

Login and Exit button

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Touch-Screen Application

During a session, the application tracks which page (side tab) you chose inside each menu

(top tab) and shows that page when you next choose that menu.

The Thermo Scientific logo icon in the upper left corner and the Login button in the lower left corner are always available.

To view the version information for the application, press the Logo icon.

Logging In to the EASY-nLC Instrument for Maintenance Tasks

To run the maintenance scripts, use the direct controls, set up the system configuration, and so on, you must log in to the EASY-nLC instrument as an administrator.

To log in as a user with administrative privileges

1. Press Login in the lower left corner of the touch screen (see Figure 19 on page 19

).

The Login As User dialog box opens (see Figure 20

).

Figure 20. Login As User dialog box

2. In the User list, select a user with administrative rights.

3. In the Password box, enter the password for this user.

Tip The factory user name and password for the system administrator is admin.

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EASY-nLC Series Troubleshooting and Maintenance Guide Thermo Scientific

4. Press Accept.

Figure 21. EASY-nLC login dialog box

1

Introduction

Touch-Screen Application

Closing Down the EASY-nLC Instrument

Closing down the instrument in a controlled manner is important to allow all of the components to shut down in an orderly sequence. Using this controlled method saves important data so that the instrument starts with the correct information the next time you use it.

IMPORTANT If you turn off the power switch during normal operation, you risk damaging essential system components.

To close down the EASY-nLC instrument and turn off the power

1. Press Exit (lower-left corner of the screen).

A confirmation dialog box opens (see

Figure 22 ).

Figure 22. Confirmation dialog box that opens after pressing Exit

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Introduction

Touch-Screen Application

2. Press Power Down.

The EASY-nLC application displays a white screen with a small progress bar. When the progress bar is filled out and the message appears indicating you can safely turn off the instrument, go to the next step.

3. After receiving the message that you can safely turn off the instrument, turn off the power switch on the back panel of the instrument.

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Specifications

Specifications

These topics describe the performance and technical specifications for the EASY-nLC syringe pumps and autosampler:

Performance Specifications

Technical Specifications

Performance Specifications

Table 3 lists the performance specifications for the EASY-nLC instruments.

Table 3. Performance specification (Sheet 1 of 2)

Item

Flow range (gradient)

Specification

20 to 2000 nL/min

Recommended: 100 to 1000 nL/min

Up to 25 μL/min Flow while loading and equilibrating

Pressure range

Retention time reproducibility (RT RSD)

EASY-nLC II: 0 to 300 bar

EASY-nLC 1000: 0 to 1000 bar

Within the recommended flow range: 0.1 to 0.4%

Outside the recommended flow range: typically better than 1%

Peak widths

Carryover

Typically 3 to 5 seconds full width at half maximum (fwhm), using PepMap™ nano-bore columns as supplied at delivery

Typically < 0.05%

Conditions: Injection of 100 fmol tryptic digests of BSA on

Pepmap columns as supplied at delivery and by using the standard wash procedure for the autosampler

Autosampler sample pickup volume range

0.1 to 18 μL with the standard 20 μL loop

0.1 to 48 μL with a 50 μL loop

Injection reproducibility

(injection RSD)

Injection linearity

0.2% at 5 μL pickup

3.0% at 0.1 μL pickup

0.9985 at 0.5 to 10 μL injection volume

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Introduction

Specifications

Table 3. Performance specification (Sheet 2 of 2)

Item

Autosampler formats

ASC model

Specification

ASA model

(earlier versions of the

EASY-nLC II instrument)

Autosampler cooling

6 × 8 Vials + 6 Vials

1 × 96 well MTP + 6 Vials

1 × 384 well MTP + 6 Vials

2 × 48 PCR strips + 6 Vials

4 × 24 PCR strips + 6 Vials

6 × 8 Vials

1 × 96 well MTP

1 × 384 well MTP

2 × 48 PCR strips

4 × 24 PCR strips

20 °C below ambient, typical setting: 7 °C (45 °F)

Technical Specifications

These tables list the technical specifications for the EASY-nLC instrument:

• General specifications, Table 4

• Physical specifications,

Table 5

• Electrical specifications,

Table 6

• Communication specifications,

Table 7

• Hardware components, Table 8

Table 4. General specifications

Item

Sound pressure level

Operating temperature

Storage temperature

Humidity

Sample viscosity

Solvent A

Specification

< 70 dBA

5 to 30 °C

–25 to +60 °C

20 to 80% RH, non-condensing

0.1 to 5 cP

Water with 0.1% formic acid

Solvent B Acetonitrile with 0.1% formic acid

Note Use only LC/MS-grade solvents

Safety According to IEC 61010

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Thermo Scientific

1

Introduction

Specifications

Table 5. Physical specifications

Item

Width

Depth

Height

Weight

Weight in the shipping container

EASY-nLC II

35 cm (13.8 in.)

38 cm (15.1 in.)

45 cm (17.7 in.)

32 kg (70.5 lb)

45 kg (99 lb)

EASY-nLC 1000

36 cm (14.2 in.)

38 cm (15.1 in.)

45 cm (17.7 in.)

35 kg (77 lb)

45 kg (99 lb)

Table 6. Electrical specifications

Item

Power requirements

Fuses

Specification

120 Vac, 50/60 Hz, 250 W

230 Vac, 50/60 Hz, 250 W

For universal power supply (UPS) dimensioning, assume 250 W.

For 120 Vac: one T 5 AL 250 V fuse (5 × 20 mm, IEC 60127)

For 230 Vac: one T 2.5 AL 250 V fuse

All fuses are UL Listed and CSA certified.

Table 7. Communication specifications

Item

Contact closure

IN circuit

OUT circuit

LAN

USB

RS-232

P-Bus

PS/2

(discontinued computer box only)

Monitor

Specification

3 outputs, 3 inputs, and 6 ground pins

TTL Level

PhotoMOS™ relay protected against high voltages. Continuous switch current from 0.8 to 5 V is 35 mA.

10/100 Mb/s BaseT Ethernet

2 × USB 1.1 for keyboard and mouse

Reserved for hardware add-ons or high-level MS control by using serial communication

Reserved for instrument add-ons, using the internal EASY-nLC instrument protocol bus:

8-wire control and limited power at 9/24 V

Input for connection of keyboard or mouse

Output for connection of external display

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Introduction

Specifications

Table 8. Hardware components

Item

Pumps

Rotary valves

Autosampler

Specification

• 140 μL volume

(enough for a >10 h, 0–100% B, 300 nL/min gradient)

• 1 nL/min to 300 μL/min flow range

• External pressure sensor

• VICI™/Valco™ rotor/stator

• 6 ports

• 3 positions (1–6, 1–2, or CENTERED)

• Peltier-cooled. Capacity is maximum 20 °C (68 °F) below ambient temperature, measured at 60% RH. Expect less if you remove the side panels, use an adapter plate in the autosampler other than the standard plate, or both.

• Plate holder ejects through spring-mounted autosampler door.

• 4 glass bottles with plastic lids for waste or wash liquids

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EASY-nLC Series Troubleshooting and Maintenance Guide Thermo Scientific

2

Maintenance Schedule

To familiarize yourself with the maintenance schedule for the EASY-nLC instrument, review this chapter.

Contents

Daily Maintenance

Weekly Maintenance

Quarterly Maintenance

Yearly Maintenance

Field Service Repairs

Daily Maintenance

Check the solvent levels, draw fresh solvent through the solvent system, and flush air out of the solvent system on a daily basis.

To check the solvent levels

1. Visually inspect solvent bottle A, solvent bottle B, and the autosampler bottle in position W3 (and the bottles in position W1 and W2 if used). Refill if necessary.

Tip You can also inspect the purity of the solvent to ensure no visible precipitates have formed that might lead to blockages in the pump line.

2. Visually inspect the autosampler wash bottle in position W4 and the plastic waste beaker in front of the solvent system compartment, and empty if necessary.

To draw fresh solvent into the system and to flush air out of the system

Run the Purge Solvent scripts with two purge iterations, and run the Flush Air script until flush volumes fall below 10 μL for the EASY-nLC II instrument or 12 μL for the

EASY-nLC 1000 instrument.

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2

Maintenance Schedule

Weekly Maintenance

Weekly Maintenance

Because the vapor pressures of formic acid, water, and acetonitrile differ, the solvent composition changes over time. Refill the solvent bottles to maintain a consistent solvent composition.

Quarterly Maintenance

Perform the following procedures quarterly (every 3 months).

To check the back pressure

To test for system blockages, run the Back Pressure script for both solvents (see “Test –

Back Pressure” on page 48 ).

IMPORTANT The script evaluates the back pressure based on using 99.9% water for solvent A and 99.9% acetonitrile for solvent B.

To check the cooler temperature

Check that the actual plate temperature matches its set point (readout on the Home >

Overview page).

Figure 23. Cooler area of Autosampler dialog box

Set point temperature

Actual temperature

To check the autosampler pickup function

Run the Sample Pickup script to check the sample pickup function (see

“Test – Sample

Pickup” on page 42 ).

To check the EASY-nLC solvent system for leaks

Run the Leaks script for “A+B” and for “System,” to check for and diagnose instrument leaks. For more information, see

“Test – Leaks” on page 44 and

“Running a System Leak

Test” on page 218 ).

IMPORTANT Check the valve rotor shifts regularly. When the valve rotor shifts exceed 6000, Thermo Fisher Scientific recommends checking the system for leaks.

From this point forward, perform leak tests every 1000 shifts and replace the rotor seal when the leak test fails.

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2

Maintenance Schedule

Yearly Maintenance

To check the check valves

Run the Purge Solvent script (see

“Prepare – Purge Solvent” on page 32

) and do the following:

• When the pumps are ejecting solvent, make sure that solvent is exiting the A and B waste tubing into the waste beaker.

• When the pumps are refilling, make sure that no solvents are going into the pump through the waste tubing.

If you detect a leaking check valve, replace it.

Yearly Maintenance

Replace the solvent filters and check the flow sensor calibration once a year.

To exchange low-pressure solvent filters and high-pressure inline filters

Thoroughly clean new solvent filters before use as these can be a contamination source.

To check the flow sensor calibration

Run the Flow Sensor calibration script with the Inspection Only parameter enabled (see

“Calibrate – Flow Sensors” on page 51 ).

This checks that the flow sensor calibration is working within specifications.

Field Service Repairs

If any of these instrument components is irreparably damaged, contact your local Thermo

Fisher Scientific field service engineer for repairs:

• Autosampler or autosampler cooler

• Rotary valve

• Pump (for both instruments) or pump PCB (for the EASY-nLC II instrument)

• Integrated computer

• Touch-screen monitor

CAUTION

Chapter 5, “Field Service Maintenance,”

describes how to replace these instrument components. However, only a Thermo Fisher Scientific field service engineer can perform these procedures. Attempted repairs by untrained personnel might cause personal injury or irreparably damage the instrument.

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Maintenance Scripts and Service Records

To familiarize yourself with the maintenance scripts and record keeping features provided by the EASY-nLC software, review this chapter.

Contents

Maintenance Scripts

Keeping Service Records

Checking and Resetting the Device Usage Counters

Maintenance Scripts

The EASY-nLC touch-screen software includes a variety of built-in maintenance scripts that help you prepare the instrument for use and troubleshoot instrument problems.

To select a maintenance script

1. On the touch screen, press Maintenance > Scripts.

2. Select a category and then the script for the specific operation you want.

You can also schedule some of the scripts for execution using the job queue.

Each of the following topics combines the category with the actual script name.

Prepare category

Prepare – Purge Solvent

Test category

Test – MS Connection

Prepare – Flush Air

Prepare – Precolumn Equilibration

Test – Sample Pickup

Test – Leaks

Prepare – Analytical Column Equilibration

Test – Valve Check

Prepare – Isocratic Flow

Test – Back Pressure

Test – Autosampler Torque

Test – Pump Torque

Calibrate category

Calibrate – Valve Tune

Calibrate – Flow Sensors

Calibrate – Reset Pressure Sensor

Calibrate – Direct Infusion

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Maintenance Scripts

Prepare – Purge Solvent

The Purge Solvent script fills the selected pump or pumps with solvent and then ejects it into the waste beaker. Use the Purge Solvent script when exchanging solvents, removing air from the solvent lines, or filling the pumps.

Figure 24 shows the parameters for the Purge Solvent script.

Figure 24. Purge Solvent script parameters

To run the Purge Solvent script

1. Open the Purge Solvent script as follows: a. On the touch screen, press Maintenance > Scripts. b. In the Category list, select Prepare. c.

In the Name list, select Purge Solvent.

2. Set up the parameters for this script as follows: a. Press the Parameters tab.

b. In the Purge Iterations box, enter the number of empty/fill cycles to be performed:

• To refresh solvents on a daily basis, enter 2 iterations.

• To exchange solvents, enter a minimum of 10 iterations.

Note Entering a value of zero (0) for purge iterations refills the pump.

c.

Select the check boxes for the appropriate pump or pumps.

3. Press Start.

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Maintenance Scripts

Prepare – Flush Air

The Flush Air script removes air from inside the pump head by pressurizing the pump and then releasing that pressure into the flow path toward the waste beaker.

The pump refills, pressurizes, releases pressure, and empties for each iteration of the script.

While the instrument executes the script, the software calculates the pressurization time dynamically based on measurements from previous iterations and measures the pumped volume required to reach 200 bar.

Note Because the solvents are slightly compressible and the pump itself expands, even an air-free pump allows some pumping before reaching 200 bar.

Depending on the solvents, a pumped volume less than 10 μL for the PLF pump model or 12 μL for the PLU pump model is acceptable.

Note

Figure 8 on page 9

shows the PLF and PLU model pumps. While the PLU model pump is standard in EASY-nLC 1000 instruments and the PLF model pump is standard in most EASY-nLC II instruments, the PLU model pump is the replacement part for both instrument models.

Figure 25 shows the parameters for the Flush Air script.

Figure 25. Flush Air script parameters

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Maintenance Scripts

To run the Flush Air script

1. Open the Flush Air script as follows: a. Press Maintenance > Scripts. b. In the Category list, select Prepare.

c.

In the Name list, select Flush Air.

2. Set up the parameters for this script as follows: a. Press the Parameters tab.

b. In the Flush Threshold [μL] box, enter the acceptable threshold for the pump model:

• For the PLF pump (standard in most EASY-nLC II instruments), enter 10.

• For the PLU pump (standard in the EASY-nLC 1000 instrument), enter 12.

3. Press Start.

The script ends after the first iteration that gives a flush volume below the set threshold.

Prepare – Precolumn Equilibration

The Precolumn Equilibration script equilibrates the precolumn with solvent from pump A.

Use this script to equilibrate or flush the precolumn and to determine a suitable flow rate for the sample loading step in your method.

Figure 26 shows the parameters for the Precolumn Equilibration script.

Figure 26. Precolumn Equilibration script parameters

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Maintenance Scripts

To run the Precolumn Equilibration script

1. Open the Precolumn Equilibration script as follows: a. Press Maintenance > Scripts. b. In the Category list, select Prepare. c.

In the Name list, select Precolumn Equilibration.

2. Set up the parameters for this script as follows: a. Press the Parameters tab.

b. Enter the appropriate values:

• In the Volume [μL] box, enter the volume of solvent A to use to equilibrate the precolumn.

For best results, use at least 10 column volumes to equilibrate the precolumn.

• In the Flow [μL/min] box, enter the flow rate for the precolumn equilibration step.

If the flow field is left empty, the pump operates at the set pressure.

• In the Max Pressure [bar] box, enter the maximum allowed pressure.

If the pressure field is left empty, the pump operates at the set flow (as long as it is below the instrument maximum pressure of 300 bar for the EASY-nLC II instrument or 1000 bar for the EASY-nLC 1000 instrument). If both a flow and a max pressure are specified, the pump flow is limited by whichever parameter is reached first.

CAUTION Check the maximum pressure rating for the precolumn. Running the instrument at pressures higher than the column’s maximum pressure rating reduces the column lifespan.

3. Press Start.

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Maintenance Scripts

Prepare – Analytical Column Equilibration

The Analytical Column Equilibration script equilibrates the analytical column with solvent from pump A.

Use this script to equilibrate or flush the analytical column, determine a suitable equilibration flow rate for the method, or initialize a new analytical column.

Figure 27 shows the parameters for the Analytical Column Equilibration script.

Figure 27. Analytical Column Equilibration script parameters

Note The Analytical Column Equilibration script sets the valves to these positions:

• Valve A: 1–6

• Valve B: Center

• Valve S: 1–2

• Valve W: 1–2

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Maintenance Scripts

To run the Analytical Column Equilibration script

1. Open the Analytical Col Equilibration script as follows: a. Press Maintenance > Scripts. b. In the Category list, select Prepare. c.

In the Name list, select Analytical Col Equilibration (abbreviated to fit in the space allotted).

2. Set up the parameters for this script as follows: a. Press the Parameters tab.

b. In the Volume [μL] box, enter the volume of solvent A to use to equilibrate the analytical column.

For best results, use at least 10 column volumes to equilibrate the analytical column.

c.

In the Flow box [μL/min], enter the flow rate for the analytical column equilibration step.

If the flow field is left empty, the pump operates at the set pressure.

d. In the Max Pressure [bar] box, enter the maximum allowed pressure.

If the pressure field is left empty, the pump operates at the set flow (as long as it is below the instrument maximum pressure of 300 bar for the EASY-nLC II instrument or 1000 bar for the EASY-nLC 1000 instrument).

If both a flow and a maximum pressure are specified, the pump flow is limited by whichever parameter is reached first.

CAUTION Check the maximum pressure rating for the analytical column.

Running the instrument at pressures higher than the maximum pressure rating for the column reduces the column lifespan.

3. Press Start.

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Maintenance Scripts

Prepare – Isocratic Flow

The Isocratic Flow script runs solvents A and B at a fixed mixture ratio. The script uses the

AFC system to accurately control the flow. Use this script to tune the mass spectrometer at a given B percentage or when cleaning the instrument, the columns, or both.

Figure 28 shows

the parameters for the Isocratic Flow script.

Figure 28. Isocratic Flow script parameters

To run the Isocratic Flow script

1. Select the Isocratic Flow script as follows: a. Press Maintenance > Scripts.

b. In the Category list, select Prepare. c.

In the Name list, select Isocratic Flow.

2. Set up the parameters (see Figure 28

) for this script as follows: a. Press the Parameters tab.

b. Enter the appropriate values:

• In the Volume box [μL], enter the total volume to be delivered by the pumps.

Note If you select the Run Indefinitely check box, the software ignores this volume setting.

• In the Flow box [μL/min], enter the flow rate to be used.

• In the AB Mix box [%B], enter the solvent composition as a percentage of solvent B.

• If you want the solvent flow to continue until you press Stop, select the Run

Indefinitely check box.

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Maintenance Scripts

3. Press Start.

When either pump runs out of solvent, both pumps refill. The refilling process takes approximately 2 minutes.

Test – MS Connection

The MS Connection script tests the contact closure function between the EASY-nLC instrument and the mass spectrometer.

Figure 29 shows the MS connection script.

Figure 29. MS Connection script

Thermo Scientific

To run the MS Connection script and test the contact closure connection

1. Connect the contact closure cable to the EASY-nLC instrument and the mass spectrometer.

Figure 30 shows the contact closure cable for an EASY-nLC instrument to be used with a

Thermo Scientific mass spectrometer.

Figure 30. Thermo Fisher contact closure cable

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Maintenance Scripts

Tip Use the Thermo Fisher contact closure cable (LC160) to make contact closure between the EASY-nLC instrument and a Thermo Scientific mass spectrometer.

Connect the two-pin connector to the mass spectrometer’s Start In pins. Connect the

12-pin connector to the back panel of the EASY-nLC instrument.

2. Check the contact closure settings for the LC/MS system as follows: a. On the EASY-nLC touch screen, press Configuration > Connections.

b. For a Thermo Scientific mass spectrometer, make the following selections:

• In the Instrument (cable no.) list, select Thermo Scientific (LC160).

• In the Protocol list under Contact Closure Settings, select One-way.

• In the State at Start list under Contact Closure Settings, select Open.

3. Turn on the mass spectrometer and set it up to wait for a contact closure signal.

• For a third-party mass spectrometer, follow the instructions in the documentation

provided with the mass spectrometer. Then, go to step 5 .

• For an LTQ Series mass spectrometer, go to

step 4

.

4. For an LTQ Series mass spectrometer, set up the mass spectrometer to wait for a contact closure signal as follows: a. Open the Tune Plus window.

b. On the menu bar, choose Control > Acquire Data.

Figure 31 shows the Control menu of the Tune Plus window for an LTQ Series mass

spectrometer.

Figure 31. Control menu

The Acquire Data dialog box opens (see

Figure 32

).

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Maintenance Scripts c.

In the Start Mode area, select the Contact Closure option.

Figure 32. Acquire Data dialog box with the Contact Closure option selected

Thermo Scientific d. Click Start.

The mass spectrometer remains in the Waiting for Contact Closure state until it receives a contact closure signal.

5. Send the contact closure signal from the EASY-nLC instrument to the MS detector as follows: a. On the EASY-nLC touch screen, press Maintenance > Scripts.

The Maintenance Scripts page opens.

b. In the Category list, select Test.

c.

In the Name list, select MS Connection.

d. Press Start.

The EASY-nLC instrument sends a contact closure signal to the mass spectrometer.

The contact closure signal triggers the mass spectrometer to start scanning. The Scan

LED on the front panel of a Thermo Scientific mass spectrometer flashes blue when the contact closure signal is set up correctly.

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Maintenance Scripts

Test – Sample Pickup

The Sample Pickup script tests the accuracy of the autosampler pickup function. During the script, the XYZ robotic arm moves to the specified sample location. Then pump S aspirates the specified volume from the vial or microplate well and ejects this volume to waste. To determine if the autosampler is aspirating the specified sample volume, weigh the sample both before and after you run the test script as described in

“Checking Sample Pickup” on page 263

.

Use this script to verify the calibration of the XYZ robotic arm and to determine the appropriate flow rate (aspiration rate by pump S) setting in the method (or Xcalibur instrument method) that you plan to use for your samples. The appropriate flow rate depends on the sample viscosity.

Figure 33 shows the parameters for the Sample Pickup script and the Position dialog box that

opens when you press the value box for the Position parameter.

Figure 33. Sample Pickup script parameters

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Maintenance Scripts

To run the Sample Pickup script

1. Load the autosampler with the appropriate samples.

Tip For more information about running this script and testing the accuracy of the

sample pickup function, see “Checking Sample Pickup” on page 263 .

2. Open the Sample Pickup script as follows: a. Press Maintenance > Scripts. b. In the Category list, select Test. c.

In the Name list, select Sample Pickup.

3. Set up the parameters for this script as follows: a. Press the Parameters tab.

b. In the Volume box [μL], enter the volume to be picked up.

c.

In the Flow box [μL/min], enter the flow rate that pump S uses to aspirate the sample. For aqueous samples enter 20 μL/min.

d. In the Position box, specify the position of the sample in the autosampler. See the

Position dialog box in Figure 33 .

4. Press Start.

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Maintenance Scripts

Test – Leaks

The Leaks script tests for possible leaks at the pump seal and in the flow path from the pump to its associated valve (high-pressure subsystems A or B). The Leaks script can also test for system leaks in the flow path downstream of valves A and B.

Use this script when you suspect leak problems in the pumps (pump piston seals), in the flow path between the pump outlet and the associated valve, or downstream of valves A and B. For information about running the system leak test, see

“Running a System Leak Test” on page 218 .

Note You cannot perform the leak test for pump S, as it does not operate under pressure during normal batch execution.

Figure 34 shows the parameters for the Leaks script

Figure 34. Leaks script parameters

During the leak test of pump A or B, the following events occur.

Step

1

2

3

4

Event

The pump refills.

The associated valve centers, blocking the solvent flow through the valve.

The pump builds pressure to 280 bar for the EASY-nLC II instrument or 980 bar for the EASY-nLC 1000 instrument.

When the pressure stabilizes at 280 bar for the EASY-nLC II instrument or 980 bar for the EASY-nLC 1000 instrument, the pump calculates the flow loss based on the pump piston movement required to maintain this pressure. If the flow loss is less than 400 nL/min, the pump passes the leak test.

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Maintenance Scripts

During a system leak test (Leaks script with System selected), the script pressurizes the flow

paths shown with red dashed lines in Figure 35 , monitors the flow sensors and pump piston

movement, and identifies the location of a leak. Before you run a system leak test, you must connect the Column Out line to the Waste In line with the HPLC union.

Figure 35. Pressurized lines for a system leak test

Pump S

Pressure sensor

Autosampler needle

Pump A

Pressure sensor

2

3

1

A

6

4 5

Flow sensor

Pump B

Pressure sensor

Mixing Tee

2

3

1

S

6

4 5

Column Out line

Waste In Line

Flow sensor

2

3

1

B

6

4 5

Centered

2

3

1

W

4 5

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Maintenance Scripts

To run the Leaks script

1. Open the Leaks script as follows: a. Press Maintenance > Scripts. b. In the Category list, select Test. c.

In the Name list, select Leaks.

2. Set up the parameters for this script as follows: a. Press the Parameters tab.

b. To select pump A or B, both pumps, or the entire system flow path for the leak test, press the cell in the Value column and make your selection from the list:

• To test the solvent path from pump A to valve A (upstream of valve A), select A.

• To test the solvent path from pump B to valve B (upstream of valve B), select B.

• To test the solvent path from pump A to valve A and from pump B to valve B

(upstream of pumps A and B), select A + B.

• To test the solvent path downstream of valves A and B, select System. Figure 35 on page 45

shows the pressurized solvent path for the system leak test.

3. For a system leak test, connect the Column Out line to the Waste In line using the union supplied in the accessory kit as shown in

Figure 35 on page 45 .

Instrument

EASY-nLC II

EASY-nLC 1000

Part number of the stainless steel union

SC600

SC900

For information about troubleshooting system leaks, see

“Troubleshooting the Results of the

System Leak Test” on page 221

and “Troubleshooting a Pump that Fails the Leaks Script” on page 216 .

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Maintenance Scripts

Test – Valve Check

The Valve Check script performs a self-test for the selected valves of an EASY-nLC II instrument.

Use this script when valves begin to move slowly or cannot reach the required positions and after exchanging valve parts (rotor or stator).

IMPORTANT This script is only valid for EASY-nLC II valves with a serial number less than V-009999. Make sure to record any numbers that the script generates. If the overshoot is greater than 6, contact your local Thermo Fisher Scientific field service engineer.

Figure 36 shows the parameters for the Valve Check script.

Figure 36. Valve Check script parameters

Thermo Scientific

To run the Valve Check script

1. Select the Valve Check script as follows: a. Press Maintenance > Scripts. b. In the Category list, select Test. c.

In the Name list, select Valve Check.

2. Set up the parameters for this script as follows: a. Press the Parameters tab.

b. Select the check boxes for the valves that you want to test.

3. Press Start.

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Maintenance Scripts

Test – Back Pressure

The Back Pressure script determines instrument back pressure for solvent A and B. The script runs at a preset flow and measures the back pressure on the system.

IMPORTANT Before you perform the Back Pressure script, ensure that the solvent A bottle contains water and the solvent B bottle contains acetonitrile. This test is not valid for other solvents.

Use this script to test for a blockage in the solvent system. Before running the script, connect the Column Out line to the Waste In line as shown in

Figure 35 on page 45 .

Figure 37 shows the parameters for the Back Pressure script.

Figure 37. Back Pressure script parameters

To run the Back Pressure script

1. Open the Back Pressure script as follows: a. Press Maintenance > Scripts. b. In the Category list, select Test. c.

In the Name list, select Back Pressure.

2. Set up the parameters for this script as follows: a. Press the Parameters tab.

b. Test solvent A, solvent B, or both A and B by selecting the corresponding check box or check boxes.

3. Press Start.

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Maintenance Scripts

Test – Autosampler Torque

The Autosampler Torque script measures the torque required to move the XYZ robot on each of its axes for the ASC autosampler.

IMPORTANT Do not run this script for the ASA autosampler. Contact your local

Thermo Fisher Scientific representative before running this script.

Figure 38 shows the Description page of the Autosampler Torque script.

Figure 38. Autosampler Torque script description

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Maintenance Scripts

Test – Pump Torque

For the PLU pump in the EASY-nLC 1000 instrument, the Pump Torque script measures the actuator’s ability to generate pressure at predefined torque levels.

IMPORTANT Do not use this script for the PLF pump in the EASY-nLC II instrument.

Contact your local Thermo Fisher Scientific representative before running this script.

Figure 39 shows the parameters for the Pump Torque script.

Figure 39. Pump Torque script parameters

Calibrate – Valve Tune

The Valve Tune script automatically retunes the selected valve in the EASY-nLC II instrument.

IMPORTANT This script is only valid for an EASY-nLC II valve with a serial number below V–009999. Contact your local Thermo Fisher Scientific representative before running this script, as running this script incorrectly will compromise the performance of the instrument.

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Maintenance Scripts

Calibrate – Flow Sensors

The Flow Sensors script calibrates the flow sensors for pumps A and B.

Use this script to do the following:

• Periodically check the accuracy of the flow sensor calibration for your maintenance records (select the Inspection Only check box).

• Calibrate the flow sensors when you change the type of solvent in solvent bottles A or B

(clear the Inspection Only check box).

This calibration does not require any additional plumbing.

IMPORTANT Because the flow sensor calibration requires a stable operating temperature, ensure that the instrument’s side panels are installed and that the instrument has been on for a minimum of one-half hour.

Figure 40 shows the parameters for the Flow Sensors script.

Figure 40. Flow Sensors script parameters

Thermo Scientific

To run the Flow Sensors script

1. Open the Flow Sensors script as follows: a. Press Maintenance > Scripts. b. In the Category list, select Calibrate. c.

In the Name list, select Flow Sensors.

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Maintenance Scripts

2. Set up the parameters for this script as follows: a. Press the Parameters tab.

b. Select the check boxes for the flow sensors (A, B, or both) that you want to either inspect or calibrate and inspect.

Note To calibrate and inspect the flow sensors, this script performs a two-point calibration as follows:

1. Calibrates the 0 μL/min point.

2. Inspects and evaluates this first calibration point.

3. Calibrates the 500 μL/min point.

4. Inspects and evaluates this second calibration point.

When you select the Inspection Only check box, the script does not apply the new calibration. Selecting the Fast Inspection option instead of the Exact

Inspection option reduces the duration of the inspection and evaluation steps from 16 minutes to 5 minutes, without reducing the accuracy of the calibration.

c.

Do one of the following:

• To check the accuracy of the current calibration (only inspect the current calibration), select the Inspection Only check box.

Note When you select the Inspection Only check box, the program does not recalibrate the flow sensors.

• To calibrate and inspect the flow sensors (and apply the new calibration), clear the Inspection Only check box.

d. Select the inspection duration as follows:

• When you want an estimate of the flow sensor accuracy, select the Inspection

Only check box and the Fast Inspection option.

• When you are recalibrating the flow sensors, clear the

Inspection Only

check box and select the Exact Inspection option.

3. Press Start.

Calibrate – Reset Pressure Sensor

This script auto-zeroes the pressure sensor.

IMPORTANT Performing this script incorrectly will compromise the performance of the instrument. Contact your local Thermo Fisher Scientific representative before running this script.

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Maintenance Scripts

Calibrate – Direct Infusion

Use the Direct Infusion script to analyze samples by direct infusion or to optimize the spray stability by infusing a known peptide.

In a direct infusion experiment, sample is introduced into the mass spectrometer without chromatographic separation by an analytical column. The EASY-nLC instrument picks up the sample volume from the specified location, and then pumps it directly into the ion source using the solvent flow from pump A.

Figure 41 shows the parameters for the Direct Infusion script.

Figure 41. Direct Infusion script

Thermo Scientific

To set up the system for a direct infusion experiment

1. Connect an emitter to the Column Out tubing.

Figure 42 shows an example of a direct infusion setup for an EASY-nLC 1000

instrument.

Figure 42. Setup for a direct infusion experiment

1/32 in. fitting Emitter and sleeve

PEEK adapter union for 1/16 in. OD and 1/32 in. OD tubing

Column Out line

1/16 in. fitting

2. Mount the assembly onto your nanoflow ion source.

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Maintenance Scripts

To run the Direct Infusion script

1. Select the Direct Infusion script as follows: a. Press Maintenance > Scripts.

b. In the Category list, select Calibrate.

c.

In the Name list, select Direct Infusion.

2. Set up the parameters for this script as follows: a. Press the Parameters tab.

b. In the Pickup Volume [μL] box, enter the volume that you want the autosampler needle to pick up. The autosampler loads this volume into the sample loop.

c.

In the Pickup Flow [μL/min] box, enter the flow rate at which pump S aspirates the sample from the vial or well.

d. In the Pickup Position box, enter the vial or well position.

e.

In Infusion Volume box [μL], enter the volume that pump A delivers to the system to backflush the sample from the sample loop out through the Column Out line and into the mass spectrometer’s ion source.

f.

In the Infusion Flow [μL/min] box, enter the flow rate at which pump A moves the sample into the mass spectrometer’s ion source.

g. Select the Wash check box to add a Wash step after pump A has pumped the specified volume of solvent A through the system.

During the Wash step, pump S empties 100 μL of solvent A through the loop and needle to the waste bottle.

3. Press Start.

IMPORTANT The Direct Infusion script flushes the solvent lines before it ends.

Stopping the script before it ends can leave sample solution in the flow path.

4. If you press Stop before the script ends, follow the instructions in message box that appears to make sure that the current sample is flushed from the system (see

Figure 43 ).

Figure 43. Wash instructions that appear when you stop the Direct Infusion script

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Keeping Service Records

Keeping Service Records

Use the Maintenance > Log Book page to electronically enter all the service actions that you perform on the instrument.

To record a service action

1. Press Maintenance > Log Book.

The Maintenance > Log Book page opens (see

Figure 44 ).

Figure 44. Log Book page of the Maintenance menu tab

Thermo Scientific

2. Press Enter Log Entry.

The Log Entry dialog box opens.

3. In the box at the top of the dialog box, enter a description of the service action.

4. In the Templates list, select a template.

The Templates list contains the following selections: Comment, New Part, Maintenance,

Property Change, and Precolumn Changed by <User> on <Date>.

5. In the Components list, select a hardware component.

The Components list contains the following selections: None, Gradient, Valve B, Valve A,

Valve S, Valve W, Analytical Column, Precolumn, Autosampler, Pump A, Pump B, and

Pump S.

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Checking and Resetting the Device Usage Counters

6. Do one of the following:

• Press Add to add this entry and keep the Log Entry dialog box open.

• Press Add and Close to add this entry and close the Log Entry dialog box.

• Press Close to close the Log Entry dialog box without saving any entries.

To search the log

1. In the Log Book list, select Queue Job or User.

2. In the User List, select Any, Admin, or Guest.

3. In the Time From list, select a time.

4. In the Time To list, select a time.

5. Press Search.

The search results appear in the Search Result table.

Checking and Resetting the Device Usage Counters

The EASY-nLC system tracks the volume pumped by pumps A, B, and S and the number of valve and rotor shifts for valves A, B, and S.

To view the device usage values

1. Press Maintenance > Devices.

2. Do one the following:

• To view the volume pumped by a pump, select the pump from the list of devices.

• To view the number of valve and rotor shifts for a valve, select the valve from the list of devices.

3. Press the Summary tab.

The Summary view opens.

To reset the volume pumped or reset the rotor shifts to zero

1. Open the appropriate Summary page.

2. Press Reset.

For a pump, the value in the Intermediate Volume box resets to 0. For a valve, the value in the Rotor Shifts box resets to 0.

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Routine Maintenance

To maintain the EASY-nLC instrument, follow the maintenance procedures in this chapter.

Tip You can access most of the instrument components by removing either the left or right side panel of the instrument. Three captive quarter-turn screws secure each panel to the instrument housing.

• To remove a side panel from the housing, loosen (unlock) the screws from the housing by rotating them 90° counterclockwise with a #2 Phillips head screwdriver. When loosened, the screws rotate freely without constraint, but remain connected to the side panel.

• To secure a side panel to the housing, mount the panel. Then tighten the screws to the housing (lock) by rotating them 90° clockwise with a #2 Phillips head screwdriver.

For information about the suggested maintenance schedule and the built-in maintenance scripts and record keeping features provided by the EASY-nLC software, see

Chapter 3,

“Maintenance Scripts and Service Records.”

Contents

Maintaining a Clean Working Environment

Replacing the Main Power Fuse

Maintaining the Syringe Pumps

Maintaining the Rotary Valves

Replacing the Check Valves

Replacing an Inline Filter for the EASY-nLC 1000 Instrument

Using nanoViper Fittings

Replacing the Autosampler Needle

Replacing the Sample Loop

Replacing a Pressure Sensor for the PLU Pump

Replacing a Flow Sensor

Replacing the Hard Drive

Managing the Devices List

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Routine Maintenance

Maintaining a Clean Working Environment

Maintaining a Clean Working Environment

Maintain a clean working environment for the EASY-nLC instrument.

When cleaning the outside of the EASY-nLC instrument, use a mild detergent and a clean cloth.

Replacing the Main Power Fuse

The EASY-nLC uses these fuses:

• For 120 Vac, the instrument uses one T 5 AL 250 V fuse (5 × 20 mm, IEC 60127).

• For 230 Vac, the instrument uses one T 2.5 AL 250 V fuse.

Use only UL Listed and CSA-certified fuses, All fuses supplied with the instrument are UL

Listed and CSA certified.

CAUTION Before removing the fuses, turn off the instrument and remove the power cable.

To replace the main power fuses

1. Move the EASY-nLC instrument to a benchtop where you can access the back panel.

Figure 45 shows the steps required to remove the fuse holder.

2. Turn off the power and pull out the power plug.

3. Place a flat-blade screwdriver into the slot in the top of the fuse holder, and then turn the screwdriver to loosen the fuse holder.

4. Pull the fuse holder out of the power entry module.

Figure 45. Replacing the main power fuse

Turn off the power and pull the power plug out of the power receptacle.

Using a flat-blade screwdriver, loosen the fuse holder.

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Routine Maintenance

Replacing the Main Power Fuse

5. To install the appropriate fuse, do one of the following:

• Insert the 5 A fuse into the slot that aligns with the white triangle at the end of the

110–120 V line power text.

• Insert the 2.5 A fuse into the slot that aligns with the white triangle at the end of the

220–240 V text.

6. Make sure that the fuse ends align with the mounting brackets.

Note The fuse holder holds two fuses. One fuse is for a 220–240 V line power and the other is for a 110–120 V line power.

7. Insert the fuse holder into the power entry module in the orientation that corresponds to the operating power.

The triangle to the right of the voltage rating points toward the white mark at the bottom of the power entry module (see

Figure 46 ).

Figure 46. Power entry module

Appropriate orientation for

220–240 line voltage

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Routine Maintenance

Maintaining the Syringe Pumps

Maintaining the Syringe Pumps

Each pump contains a piston seal and a piston. Over time, the buffered solutions leave deposits on the pistons, and through constant contact with the moving pistons, the piston seals slowly deteriorate.

Note Most EASY-nLC II instruments contain PLF model pumps, which have been discontinued. If this pump is damaged, you must replace the PLF model pump with a

PLU model pump.

Both the EASY-nLC 1000 instrument and new shipments of the EASY-nLC II instrument use PLU model pumps.

Figure 8 on page 9 shows the two pump models.

To replace a pump piston seal and clean the piston, follow these procedures:

1.

“Retracting the Piston” on page 61

2. Depending on the pump model, follow one of these procedures:

“Replacing the Piston Seal and Cleaning the Piston in a PLF Pump” on page 62

“Replacing the Piston Seal in a PLU Pump” on page 67

3.

“Priming the Pump” on page 71

4.

“Resetting the Pump Usage Counter” on page 72

5.

“Removing Air After Replacing a Piston Seal or a Pump” on page 73

6. Depending on whether you are performing maintenance on one of the chromatography pumps (pump A or B) or on the sample pump (pump S), follow one of these procedures:

• For pumps A and B, run a pump leak test as described in “Running the Leaks Script after Replacing a Piston Seal or a Pump” on page 73 .

• For pump S, run a sample pickup test as described in “Test – Sample Pickup” on page 42 .

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Routine Maintenance

Maintaining the Syringe Pumps

Retracting the Piston

Set the pump piston to the fully retracted position by running the Purge Solvent script with 0 iterations.

To retract the pump piston

1. On the touch screen, press Maintenance > Scripts.

2. In the Category list, select Prepare.

3. In the Name list, select Purge Solvent.

4. Press the Parameters tab.

5. In the Purge Iterations box, enter 0 (see Figure 47

).

Figure 47. Purge solvent script with 0 iterations

Thermo Scientific

6. Select the check box for the appropriate pump.

7. Press Start.

The piston moves backward until it reaches the 140 μL position.

8. To replace the piston seal, go to the piston seal replacement procedure for your pump.

“Replacing the Piston Seal and Cleaning the Piston in a PLF Pump” on page 62

–or–

“Replacing the Piston Seal in a PLU Pump” on page 67

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Routine Maintenance

Maintaining the Syringe Pumps

Replacing the Piston Seal and Cleaning the Piston in a PLF Pump

Most EASY-nLC II instruments have PLF model pumps (see Figure 8 on page 9

).

Note To replace the piston seal in a PLU pump, follow the instructions in

“Replacing the

Piston Seal in a PLU Pump” on page 67

.

Replacing a piston seal in the PLF pump requires these tools and materials.

Tools

• #2 Phillips head screwdriver

• 13 mm open-ended wrench

• 1/4 in. open-ended wrench

• Pipette

Parts and materials

• Powder-free safety gloves

• LC/MS-grade methanol

• LC/MS-grade acetonitrile

• Piston seal and piston seal tool, P/N LC210

CAUTION Wear powder-free gloves and safety glasses when handling parts of the LC system that come into contact with solvents.

To replace the piston seal in a PLF model pump

1. If you have not already done so, set the piston to its fully retracted position as described in

“Retracting the Piston” on page 61

.

2. Set the corresponding valve to the Center position as described in “Using the Valve

Controls” on page 204 .

3. Close down the EASY-nLC instrument and turn off the power (see “Closing Down the

EASY-nLC Instrument” on page 21 ).

4. Using a #2 Phillips head screwdriver, with a quarter-turn loosen the three captive screws that secure the right side panel to the instrument housing. Then remove the panel.

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Maintaining the Syringe Pumps

5. Remove the tubing connected to the pump head as follows:

• If a PEEK fitting is connected to the pump head, use a 13 mm open-ended wrench, to remove it (see

Figure 48

).

Figure 48. Removing a PEEK fitting from the externally threaded pump head for a PLF model pump

PEEK fitting connected to an externally threaded pump head

• If a stainless steel fitting is connected to the pump head, use a 1/4 in. open-ended wrench to remove it.

Figure 49 shows a stainless steel fitting connected to an internally threaded pump

head.

Figure 49. Internally threaded pump head for a PLF model pump

Stainless steel fitting connected to an internally threaded pump head

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Routine Maintenance

Maintaining the Syringe Pumps

6. Using a 13 mm open-ended wrench, remove the pump head (see

Figure 50

).

Figure 50. Removing the pump head

Pump head (PLF model pump)

7. Clean the piston as follows: a. Place the piston in the fully extended position as follows: i.

Turn on the instrument and log in as an administrator.

ii. Press Home > Overview. iii. Press the pump icon for the pump you want to control.

The Pump dialog box opens.

iv. In the flow rate box, enter the flow rate: 300 μL/min. v.

In the volume box, enter the dispense volume: 140 μL.

vi. Press Start.

b. Soak a lint-free tissue in methanol, and then squeeze out the excess solvent.

c.

Clean the piston with the lint-free tissue soaked in methanol, and visually inspect the piston for any scratches. Take care to avoid solvent drips, as the pump printed circuit board can be damaged by exposure to solvents.

CAUTION The pump PCB is easily damaged by exposure to solvents.

d. Return the piston to the fully retracted position as follows: i.

In the Pump dialog box, set the flow rate to – 300 μL/min and the dispense volume to 140 μL. ii. Press Start.

8. When the piston is fully retracted, close down the EASY-nLC system and turn off the power (see

“Closing Down the EASY-nLC Instrument” on page 21

).

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Routine Maintenance

Maintaining the Syringe Pumps

9. Remove the worn piston seal as follows: a. To avoid contaminating the pump head, clean the piston seal tool and guide tool by soaking them in a beaker filled with 100% HPLC-grade methanol.

Figure 51 shows the piston seal tool.

Figure 51. Piston seal tool for the PLF model pump

Piston seal tool

Guide tool b. Insert the piston seal tool into the worn piston seal and pull the piston seal out of the pump head (

Figure 52

).

Figure 52. Using the piston seal tool to remove the worn piston seal (PLF model pump)

Worn piston seal

10. Install the new piston seal as follows: a. Insert the guide tube into the pump head flange (see

Figure 53 ).

Figure 53. Guide tool inserted into the pump head flange (PLF model pump)

Guide tool

Thermo Scientific b. Using a pipette, fill the groove in the piston seal with methanol.

Tip Filling the piston seal groove with methanol before installing the seal reduces the time required for the Flush Air script to remove air from the pump.

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Routine Maintenance

Maintaining the Syringe Pumps c.

Place the new seal with the groove facing upward on the piston seal tool

(see

Figure 54 ).

Figure 54. Piston seal mounted onto the piston seal tool (PLF model pump) d. Insert the piston seal tool into the guide tool (see

Figure 55 ).

Figure 55. Inserting the piston seal tool into the guide (PLF model pump) e.

Push the piston seal tool into the guide until you feel resistance (see

Figure 56

).

Figure 56. Seating the piston seal (PLF model pump)

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Routine Maintenance

Maintaining the Syringe Pumps f.

Remove the guide tube and the piston seal tool. and then check that the piston seal is properly seated (see

Figure 57 ).

Figure 57. Piston seal properly seated in the pump head (PLF model pump)

11. Using a 13 mm open-ended wrench, reconnect the pump head to the pump body.

12. Go to “Priming the Pump” on page 71 .

Replacing the Piston Seal in a PLU Pump

Replacing the piston seal in a PLU model pump requires these tools and materials.

Tools

• #2 Phillips head screwdriver

• 3 mm L-hex wrench

• 1/4 in. open-ended wrench

• Pipette

Parts and materials

• Powder-free safety gloves

• LC/MS-grade methanol

• LC/MS-grade acetonitrile

• Piston seal and piston seal tool, P/N LC510

To replace the piston seal in a PLU model pump

1. If you have not already done so, set the piston to its fully retracted position as described in

“Retracting the Piston” on page 61

.

2. Set the corresponding valve to the Center position as described in “Using the Valve

Controls” on page 204 .

3. Close down the EASY-nLC instrument and turn off the power (see “Closing Down the

EASY-nLC Instrument” on page 21 ).

4. Using a #2 Phillips head screwdriver, with a quarter-turn loosen the three captive screws that secure the right side panel to the instrument housing. Then remove the panel.

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Routine Maintenance

Maintaining the Syringe Pumps

5. Remove the used piston seal as follows: a. Using a 1/4 in. open-ended wrench, remove the stainless steel fitting connected to the

pump head (see Figure 58 ).

Figure 58. Stainless steel fitting connected to the pump head (PLU model pump)

Stainless steel fitting b. Using a 3 mm L-hex wrench, remove the two screws that secure the pump head to

the pump body (see Figure 59 ).

IMPORTANT Do not move the piston if the pump head has been removed.

Doing so can affect the piston alignment, which might cause the piston calibration to be lost and result in irreversible piston damage.

Figure 59. Using a 3 mm L-hex wrench to remove the pump head

Two screws that secure the pump head to the pump body c.

Clean the exposed portion of the piston with a lint-free tissue soaked in HPLC-grade methanol and visually inspect the piston for scratches. Ensure that no solvent runs into the pump.

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4

Routine Maintenance

Maintaining the Syringe Pumps d. Using only your fingernails or a plastic tool, remove the old seal from the pump head

(see

Figure 60 ).

CAUTION Avoid using a metal tool to remove the piston seal from the pump head, as metal can scratch the inside of the pump head and generate leaks.

Figure 60. Worn piston seal installed in the pump head (PLU model pump)

6. Install the new piston seal as follows: a. With the spring side of the piston seal facing upward, mount the new piston seal onto the piston seal tool (see

Figure 61

).

Figure 61. Piston seal mounted on the piston seal tool (PLU model pump)

Piston seal spring b. Using a pipette, fill the rim of the piston seal with 100% LC/MS-grade methanol to

remove the air from the piston seal spring (see Figure 62 ).

Figure 62. Filling the piston seal with methanol (PLU model pump)

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Routine Maintenance

Maintaining the Syringe Pumps c.

Place the pump head on the guide rods and evenly press it all the way down against

the piston seal tool to insert the seal into the pump head (see Figure 63 ).

Figure 63. Inserting the piston seal into the pump head (PLU model pump)

d. Gently pull the pump head off of the guide rods (see Figure 64

).

Figure 64. Removing the pump head from the piston seal tool (PLU model pump)

7. Using the two screws and a 3 mm L-hex wrench, reconnect the pump head to the pump body. Do not connect the stainless steel tubing to the pump head.

8. Go to the next procedure,

“Priming the Pump.”

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Maintaining the Syringe Pumps

Priming the Pump

After you replace a piston seal, prime the pump with the appropriate solvent before you reconnect the solvent line to the pump head.

To prime the pump

1. Turn on the EASY-nLC instrument and log in as an administrator.

2. Set the piston to its fully extended position as follows: a. On the touch screen, press Home > Overview.

b. Press the pump icon for the pump that you want to control.

The Pump X dialog box opens, where X identifies the pump (A, B, or S).

c.

Set the flow rate to 300 μL/min and the volume to 140 μL. d. Press Start.

3. Fill the pump head with solvent as follows:

a. Pipette an aliquot of the appropriate solvent into the pump head (see Figure 65

).

Figure 65. Adding solvent to the pump head

Thermo Scientific

CAUTION Be careful not to spill solvent on the PCBs.

• For the PLU pump model (standard in the EASY-nLC 1000 instrument and replacement part for both EASY-nLC instruments), the LED panel cover for each pump protects the PCB from accidental contact when the instrument’s right panel is removed. However, the cover does not form a waterproof seal with the pump body so that the PCBs are exposed to solvent leaks from above.

• For the PLF pump model (EASY-nLC II instrument), the printed circuit boards are completely exposed.

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Routine Maintenance

Maintaining the Syringe Pumps b. In the Pump dialog box, set the flow rate to –300 μL/min and the volume to 140 μL.

Then press Start.

The piston moves down, drawing the solvent into the pump. c.

Make sure the pump head is filled with solvent during the entire retraction of the piston.

4. Reconnect the solvent line to the pump head as follows:

• For a PLF pump with a PEEK fitting (see

Figure 48 on page 63

), use a 13 mm open-ended wrench to tighten the fitting.

• For a PLF pump with a stainless steel fitting (see

Figure 49 on page 63 ) or a

PLU pump, use a 1/4 in. open-ended wrench to tighten the fitting.

5. Mount the right side panel to the instrument housing. Then, with a #2 Phillips head screwdriver, secure the panel by rotating the three captive screws a quarter-turn clockwise.

After you reconnect the pump head to the pump body and the high-pressure line from the pressure sensor to the pump head, reset the pump usage counter, and then remove air from the system.

Note You reset the usage counters before removing air from the system because the air removal procedure requires the pump to go through 10 purge cycles.

Resetting the Pump Usage Counter

Reset the pump usage counter after you prime the pump.

To reset the pump usage counter

1. On the touch screen, press Maintenance > Devices.

2. Select the pump from the list of devices.

3. Press the Summary tab.

4. Press Reset.

The value in the Intermediate Volume box resets to 0.

For more information, see “Checking and Resetting the Device Usage Counters” on page 56 .

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Routine Maintenance

Maintaining the Syringe Pumps

Removing Air After Replacing a Piston Seal or a Pump

To draw fresh solvent through the solvent system and to remove air from the solvent system components, including the pump heads, follow these procedures after you prime the pump and reset the usage counters.

To draw fresh solvent into the system and remove air from the system

1. To draw fresh solvent into the system and remove air from the solvent lines, do the following: a. On the touch screen, press Maintenance > Scripts.

b. In the Category list, select Prepare.

c.

In the Name list, select Purge Solvent (see

“Prepare – Purge Solvent” on page 32 ).

d. Press the Parameters tab.

e.

In the Purge Iterations box, enter 10.

f.

Select the check box for the appropriate pump. g. Press Start and wait for the system to perform 10 purge cycles.

2. To remove air from the pump head, do the following: a. In the Name list, select Flush Air (see

“Prepare – Flush Air” on page 33 ).

b. Press the Parameters tab. c.

Set the flush volume threshold to 10 μL for the PLF pump or 12 μL for the PLU pump.

Note

Figure 8 on page 9

shows the PLU and PLF pump models.

d. Press Start, and wait for the script to finish.

Running the Leaks Script after Replacing a Piston Seal or a Pump

After you prime pump A or pump B, reset the usage counters, remove air from the system, and run a pump leak test.

Note For pump S, run a sample pickup test as described in “Test – Sample Pickup” on page 42

.

For information about replacing a pump, see “Replacing a Pump” on page 150 . Only a

Thermo Fisher Scientific field service engineer can replace a damaged pump, as replacing the pump is an advanced maintenance procedure.

To run a pump leak test on a replacement pump or a pump with a new piston seal

Follow the instructions in

“Test – Leaks” on page 44 . When the instrument is leak tight,

the instrument is ready for use.

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Routine Maintenance

Maintaining the Rotary Valves

Maintaining the Rotary Valves

The EASY-nLC instrument has four rotary valves labeled Valve A, Valve B, Valve S, and

Valve W. These valves contain the same internal components.

To determine if the valve requires maintenance, run the Leak test script as follows:

• For valves S and W, run the Leaks script on the system (System).

• For valves A and B, run the Leaks script on the respective subsystem (A or B).

To maintain the valves, follow these procedures:

“Cleaning the Rotor Seal and Stator,” on this page

“Replacing the Rotor Seal” on page 77

“Replacing the Stator” on page 78

Maintaining the valves by cleaning the rotor seal and stator or by replacing the rotor seal, the stator, or both parts requires these tools and materials.

Tools

• 9/64 in. hex wrench

• 1/4 in. open-ended wrench

• #2 Phillips head screwdriver

Materials

• Powder-free safety gloves

• LC/MS-grade methanol

• Lint-free cloth

CAUTION Wear powder-free gloves and safety glasses when handling parts of the LC system that come into contact with solvents.

Cleaning the Rotor Seal and Stator

If a rotary valve is leaking, attempt to remove the leak by cleaning the rotor seal and the stator before taking the more costly step of replacing the rotor seal.

To clean the rotor seal and stator in a valve and return the instrument to normal operation, follow these steps:

1.

“To remove the stator from the valve” on page 75

2.

“To clean the rotor seal and the stator” on page 75

3.

“To reassemble the cleaned valve and return the instrument to normal operation” on page 76

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Routine Maintenance

Maintaining the Rotary Valves

To remove the stator from the valve

1. Close down the EASY-nLC instrument, and then turn off the power to the instrument

(see

“Closing Down the EASY-nLC Instrument” on page 21 ).

2. Using a #2 Phillips head screwdriver, with a quarter-turn loosen the three captive screws that secure the right side panel to the instrument housing. Then remove the panel.

3. Using a 9/64 in. L-hex wrench, remove the two hex screws that secure the stator to the valve assembly, and then pull the stator away from the valve driver.

4. Depending on whether you are cleaning or replacing the rotor seal, go to the appropriate procedure:

“To clean the rotor seal and the stator,”

on this page

“To replace the rotor seal” on page 77

Note If you are cleaning the valve components or replacing only the rotor seal, you do not need to remove the tubing connected to the stator. If you are replacing the stator, you must remove all the solvent lines from the valve.

To clean the rotor seal and the stator

1. If you have not already done so, remove the stator from the valve as described in the

previous procedure, “To remove the stator from the valve.”

2. Leave the rotor seal mounted to the valve driver (see

Figure 66 ). Clean the mounted rotor

seal with a lint-free tissue or Q-tip soaked in methanol.

Figure 66. Rotor seal mounted on the valve driver

Rotor seal

3. Clean the stator with a lint-free tissue or Q-tip soaked in methanol (see

Figure 67

).

Figure 67. Cleaning the stator

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Maintaining the Rotary Valves

To reassemble the cleaned valve and return the instrument to normal operation

1. Mount the clean stator onto the valve driver.

2. Using a 9/64 in. L-hex wrench, tighten the two hex screws a little at a time by shifting from one screw to the other and back again until the screws are evenly torqued.

3. Mount the right side panel to the instrument housing. Then, with a #2 Phillips head screwdriver, secure the panel by rotating the three captive screws a quarter-turn clockwise.

4. Turn on the EASY-nLC instrument and log in as an administrator or a super user.

5. Go to the next procedure,

“To remove air from the system and check for leaks after disassembling a valve.”

To remove air from the system and check for leaks after disassembling a valve

1. If you removed the stator from valves A or B, flush air from the system as follows: a. Set up the Flush Air script as described in

“Prepare – Flush Air” on page 33 .

b. Select the pump that is connected to the affected valve.

c.

Set the flush volume threshold to 10 μL for the EASY-nLC II or 12 μL for the

EASY-nLC 1000.

d. Press Start.

e.

Wait for the script to finish.

2. Check the system or subsystem for leaks as follows:

• If you removed the stator from in valve A or B, run the Leaks script for the affected

valve (see “Test – Leaks” on page 44

).

• If you replaced the rotor seal in valve S or W, run the Leaks script for the system.

When the system is leak tight, the instrument is ready for use.

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Routine Maintenance

Maintaining the Rotary Valves

Replacing the Rotor Seal

If cleaning the rotor seal and stator does not remove the system leak, replace the rotor seal.

Replacing the rotor seal requires one of these parts.

Valve serial number

V-010000 and above

V-009999

Replacement part

Rotor seal, P/N LC228

Rotor seal, P/N LC224

Tip The About page of the Rotary Valve view on the Maintenance > Devices page lists the serial number of the selected valve.

To replace the rotor seal in a valve and return the instrument to normal operation, follow these steps:

1.

“To remove the stator from the valve” on page 75

2.

“To replace the rotor seal,” on this page

3.

“To reset the usage counter for the valve” on page 78

4.

“To remove air from the system and check for leaks after disassembling a valve” on page 76

To replace the rotor seal

1. If you have not already done so, access the rotor seal by following “To remove the stator from the valve” on page 75

.

2. Remove the rotor seal from the valve driver (see Figure 68 ).

Figure 68. Rotor seal mounted on the valve driver

Rotor seal

3. Carefully mount the new rotor onto the valve driver, ensuring that the rotor sealing surface (engraved flow passage), is facing out.

4. If you have not already done so, clean the stator with a lint-free tissue or Q-tip soaked in methanol.

5. Mount the clean stator onto the valve driver.

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Maintaining the Rotary Valves

6. Using a 9/64 in. L-hex wrench, tighten the two hex screws a little at a time by shifting from one screw to the other and back again until the screws are evenly torqued.

7. Mount the right side panel to the instrument housing. Then, with a #2 Phillips head screwdriver, secure the panel by rotating the three captive screws a quarter-turn clockwise.

To reset the usage counter for the valve

1. Turn on the EASY-nLC instrument and log in to the system as an administrator.

2. Reset the usage counter for the valve as follows: a. Press Maintenance > Devices. b. Select the valve from the list of devices.

c.

Press the Summary tab.

d. Press Reset.

The value in the Rotor Shifts box resets to 0.

For more information, see “Checking and Resetting the Device Usage Counters” on page 56 .

3. To remove air from the system and check for leaks, follow “To remove air from the system and check for leaks after disassembling a valve” on page 76 .

Replacing the Stator

Replacing the stator requires one of these parts.

Valve serial number

V-001000–V-099999

V-000100–V-000999

Replacement part

Coated stator, P/N LC226

Uncoated stator, P/N LC225

Tip The About page of the Rotary Valve view on the Maintenance > Devices page lists the serial number of the selected valve.

To replace the stator in a valve and return the instrument to normal operation, follow these steps:

1.

“To remove the stator from the valve assembly” on page 79

2.

“To install the new stator” on page 79

3.

“To remove air from the system and check for leaks after disassembling a valve” on page 76

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Maintaining the Rotary Valves

To remove the stator from the valve assembly

1. Close down the EASY-nLC system, and then turn off the power to the instrument

(see

“Closing Down the EASY-nLC Instrument” on page 21 ).

2. Using a #2 Phillips head screwdriver, with a quarter-turn loosen the three captive screws that secure the right side panel to the instrument housing. Then remove the panel.

3. Disconnect the solvent lines from the valve as follows:

• Use a 1/4 in. open-ended wrench to remove stainless steel fittings.

• Use the black nanoViper knurled nut to loosen and remove nanoViper fittings.

4. Using a 9/64 in. L-hex wrench, remove the two hex screws that secure the stator to the valve driver, and then pull the stator away from the valve driver.

To install the new stator

1. Mount the new stator onto the valve driver.

2. Using a 9/64 in. L-hex wrench, tighten the two hex screws a little at a time by shifting from one screw to the other and back again until the screws are evenly torqued.

3. Reconnect the solvent lines to the valve as follows:

• For stainless steel fittings, use a 1/4 in. open-ended wrench to tighten the fittings.

• For the nanoViper fittings in an EASY-nLC 1000 instrument, follow the instructions

in “Using nanoViper Fittings” on page 83 to reconnect the nanoViper solvent lines to

the valve ports. Take care not to overtighten these fingertight fittings.

4. To remove air from the system and check for leak, follow “To remove air from the system and check for leaks after disassembling a valve” on page 76 .

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Routine Maintenance

Replacing the Check Valves

Replacing the Check Valves

If a check valve is not functioning, replace it.

Replacing a check valve requires the following tools and materials.

Tools

• 9/16 in. open-ended wrench

• #2 Phillips head screwdriver

Parts and materials

• Powder-free safety gloves

• Solvent-side check valve, P/N LC233

• Waste-side check valve, P/N LC234

CAUTION Wear powder-free gloves and safety glasses when handling parts of the LC system that come into contact with solvents.

To replace a check valve

1. Close down the EASY-nLC system, and then turn off the power to the instrument

(see

“Closing Down the EASY-nLC Instrument” on page 21 ).

2. Using a #2 Phillips head screwdriver, with a quarter-turn loosen the three captive screws that secure the right side panel to the instrument housing. Then remove the panel.

3. Do the following:

• For a solvent-side check valve, remove the solvent inlet line from the check valve, and then terminate the solvent line with a plug.

• For a waste-side check valve, remove the waste line from the check valve.

CAUTION To prevent solvent from siphoning out of the solvent bottle on top of the instrument and onto system components, use a plug to terminate a solvent inlet line when you disconnect it from the solvent-side check valve.

The pump PCB is easily damaged by contact with solvents.

4. Do one of the following:

• For a solvent-side check valve, use a 9/16 in. open-ended wrench to loosen the valve from the check valve assembly.

• For the waste-side check valve, remove the check valve by turning it counterclockwise with your fingers.

5. Screw the new check valve onto the check valve assembly. Using a 9/16 in. open-ended wrench, slightly tighten the solvent-side check valve to the assembly.

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Routine Maintenance

Replacing an Inline Filter for the EASY-nLC 1000 Instrument

6. Do one of the following:

• For a solvent-side check valve, reconnect the solvent inlet line.

• For a waste-side check valve, reconnect the waste line.

7. Mount the right side panel to the instrument housing. Then, with a #2 Phillips head screwdriver, secure the panel by rotating the three captive screws a quarter-turn clockwise.

8. Run the Purge Solvent script for the pump associated with the replaced check valve, and monitor the flow from the solvent bottle when the pump is filling and the flow to the waste beaker when the pump is emptying (see

“Prepare – Purge Solvent” on page 32 ).

Replacing an Inline Filter for the EASY-nLC 1000 Instrument

Inline filters are connected to both the inlet and outlet ports of the flow sensors. The inline filter consists of a stainless steel body (nut, filter, and tube) and a ferrule (see

Figure 69 ).

Figure 69. Inline filter components (nut with integrated tube and filter and ferrule)

Thermo Scientific

Replacing an inline filter requires the following tools and materials.

Tools

• 5/16 in. open-ended wrench

• #2 Phillips head screwdriver

Parts and materials

• Powder-free safety gloves

• Inline filter

CAUTION Wear powder-free gloves and safety glasses when handling parts of the LC system that come into contact with solvents.

To install an inline filter into the receiving port of a flow sensor

1. Close down the EASY-nLC system, and then turn off the power to the instrument

(see

“Closing Down the EASY-nLC Instrument” on page 21 ).

2. Using a #2 Phillips head screwdriver, with a quarter-turn loosen the three captive screws that secure the right side panel to the instrument housing. Then remove the panel.

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Replacing an Inline Filter for the EASY-nLC 1000 Instrument

3. While holding the wide end of the ferrule toward the nut, slide the ferrule over the tube that extends from the threaded end of the nut.

IMPORTANT The inline filters have a swept volume (flow path volume) of less than one microliter.

To prevent leaks, take care to properly seat the ferrule in the receiving port. Once the ferrule is swaged onto the tubing, do not connect the fitting to a different receiving port.

4. Using a 5/16 in. open-ended wrench, tighten the nut.

5. Install the nanoViper solvent inlet line into the inline filter (see

Figure 70 ).

Figure 70. Installed inline filters

Inline filter

6. Run the Back Pressure script and the Leak script on the system as described in these topics:

“Test – Back Pressure” on page 48

“Test – Leaks” on page 44

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Routine Maintenance

Using nanoViper Fittings

Using nanoViper Fittings

For the EASY-nLC instrument, most of the plumbing connections are made with nanoViper fittings (see

Figure 71

).

Note For more information about working with nanoViper fittings, see

“Using nanoViper Fittings Quick Reference Guide” on page 323 .

Figure 71. nanoViper fitting

PEEK sealing surface

Removable, black knurled tightening tool

Thermo Scientific

Screw

Even though nanoViper fittings can withstand UHPLC backpressures of up to ~1034 bar

(~15 000 psi), they are fingertight fittings, which require only very small torques to seal.

Therefore, you must follow the guidelines below to avoid damage by overtightening.

To use a nanoViper fitting

1. Insert the nanoViper fitting into the target port and slowly rotate the screw clockwise until you feel resistance.

2. Using the black knurled tightening tool, turn the screw clockwise to an angle between

0 and 45 degrees (1/8-turn).

3. Run the Leaks script for the system as described in

“Running a System Leak Test” on page 218 .

When the leak test ends, the system is at atmospheric pressure.

IMPORTANT To extend the lifetime of the nanoViper fittings, open and close connections at atmospheric system pressures only. Opening and closing connections at high system pressures can reduce the lifetime of the fitting system.

4. If the Leaks script fails because the new connection is not leak tight, use the black knurled tightening tool to turn the screw by as much as an additional 45 degrees. Do not turn the screw beyond an angle of 90 degrees from where you felt the initial resistance.

IMPORTANT To prevent damage to the sealing surface of the nanoViper fitting, take care not to overtighten the nanoViper fitting.

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Routine Maintenance

Replacing the Autosampler Needle

Replacing the Autosampler Needle

Replacing the autosampler needle requires these tools and materials.

Tools

• 1/4 in. open-ended wrench

• #2 Phillips head screwdriver

Parts and materials

• Powder-free safety gloves

• ASA autosampler needle, P/N LC251

• ASC autosampler needle, P/N LC302

To remove the autosampler needle from an ASC or an ASA model autosampler

1. Using a #2 Phillips head screwdriver, with a quarter-turn loosen the three captive screws that secure the right side panel to the instrument housing. Then remove the panel.

2. Using a 1/4 in. open-ended wrench, unscrew the nut that secures the autosampler needle to port 1 of valve S. Then remove the fitting from the port.

Figure 72 and Figure 73

show the solvent line connections to valve S for the

EASY-nLC II solvent system and the EASY-nLC 1000 solvent system, respectively.

Figure 72. Valve S solvent line connections for the EASY-nLC II instrument

Valve S

1

1

Autosampler needle

2

2

6

6

3

3

5

5

4

4

From the pressure sensor

Column Out line

From the mixing Tee

Figure 73. Valve S solvent line connections for the EASY-nLC 1000 instrument

Valve S

1

2

6

Autosampler needle

3

5

From the pressure sensor

4

Column Out line

From the mixing Tee

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Replacing the Autosampler Needle

3. Remove the nut, ferrule, and sleeve from the needle tubing.

Figure 74. Autosampler tubing with a nut, ferrule, and sleeve at one end

4. Depending on the instrument model, move the z-axis needle holder to an accessible location within the tray compartment by doing the following:

• For the ASA model, go to

step 5

.

• For the ASC model, go to

step 6

.

5. For the ASA model, move the z-axis needle holder to position A1 as follows: a. Press Home > Overview.

b. Press the Autosampler icon.

The Autosampler dialog box opens.

c.

In the XYZ Robot area, select position A1 in the Well box (see Figure 75 ).

Figure 75. Autosampler direct control dialog box

Thermo Scientific d. Press Goto.

e.

Go to step 7 on page 86 .

6. For the ASC model, move the z-axis needle holder to position W1 as follows: a. Press Home > Overview. b. Press the Autosampler icon.

The Autosampler dialog box opens.

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Routine Maintenance

Replacing the Autosampler Needle c.

In the XYZ Robot area, press Go to Calibration.

The Tools page of the Autosampler view on the Maintenance > Devices page opens

(see

Figure 76 ).

Figure 76. Tools page of the Autosampler view on the Maintenance > Devices page d. In the Manipulator area, press Reset.

The z-axis needle holder moves to the W1 position.

7. Using a #2 Phillips head screwdriver, with a quarter-turn loosen the three captive screws that secure the right side panel to the instrument housing. Then remove the panel.

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Routine Maintenance

Replacing the Autosampler Needle

8. Depending on the instrument model, do the following:

• For the ASC model, remove the black needle holder from the slot in the metal plate that separates the solvent system hardware from the autosampler compartment.

Removing the fitting requires some gentle movement from both sides of the plate

(see

Figure 77 ).

Figure 77. Removing the black needle holder from the slot in the metal plate

Black needle holder Slot in the metal plate

• For the ASA model, remove the white plastic needle guide on the back of the touch-screen monitor.

9. Loosen the nut that is connected to the z-axis needle holder by turning it

counterclockwise (see Figure 78 ). Then carefully pull the autosampler needle upward and

away from the holder.

Figure 78. Autosampler needle connected to the z-axis needle holder

ASA model z-axis needle holder ASC model z-axis needle holder

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Routine Maintenance

Replacing the Autosampler Needle

To install a new autosampler needle

1. Mount the new autosampler needle in the z-axis holder. Check that the small spring is situated between the PEEK nut and the plastic stop.

2. Do one of the following:

• For the ASC autosampler, go to step

step 3

.

• For the ASA autosampler, go to step

step 4

.

3. For the ASC autosampler, do the following: a. Guide the end of the needle that connects to valve S through the large hole in the metal plate that separates the tray compartment from the solvent system compartment. b. Install the black needle holder in the side plate.

4. For the ASA autosampler, slide the valve end of the tubing through the small plastic holder on the back of the touch-screen monitor (see

Figure 79 ).

Figure 79. Holder on the back of the touch-screen monitor

Holder

5. Connect the needle to port 1 of valve S as follows: a. Slide the provided blue sleeve and metal fittings onto the tubing. b. To ensure a zero dead volume connection, hold the blue sleeve and the tubing firmly against the bottom of the valve port, and then tighten the fitting with a

1/4 in. open-ended wrench.

6. Recalibrate the needle position as described in Chapter 7, “Calibrating the Autosampler’s

XYZ Robot.”

7. Run 2 iterations of the Purge Solvent script for Pump S.

For information about running the Purge Solvent script, see “Prepare – Purge Solvent” on page 32 .

8. Run the Flush Air script for Pump S with a flush volume threshold of 10 μL for the

EASY-nLC II system or 12 μL for the EASY-nLC 1000 system.

For information about running the Flush Air script, see “Prepare – Flush Air” on page 33 .

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Replacing the Sample Loop

Replacing the Sample Loop

Thermo Fisher Scientific provides sample loops in several sizes for the EASY-nLC instruments.

To replace the sample loop

1. Using a #2 Phillips head screwdriver, with a quarter-turn loosen the three captive screws that secure the right side panel to the instrument housing. Then remove the panel.

2. To remove the sample loop from the valve, do one of the following:

• For the EASY-nLC II instrument, use a 1/4 in. open-ended wrench to loosen the fittings that secure the sample loop to ports 2 and 5 of valve S. Then remove the sample loop from the valve.

Figure 80 shows the sample loop connections for the EASY-nLC II instrument.

Figure 80. Sample loop connections for the EASY-nLC II instrument

Sample loop

2

2

3

3

1

1

4

4

5

6

6

5

• For the EASY-nLC 1000 instrument, do the following:

a. Return the system to atmospheric pressure (see “Returning the System to

Atmospheric Pressure” on page 325

). b. Disconnect the nanoViper fittings connected to ports 2 and 5 of valve S.

Figure 81 shows the sample loop connections for the EASY-nLC 1000 instrument.

Figure 81. Sample loop connections for the EASY-nLC 1000 instrument

1

Sample loop

2

6

5

3

4

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Replacing a Pressure Sensor for the PLU Pump

3. Connect the replacement sample loop to valve S as follows: a. Insert the fittings at the end of the sample loop into ports 2 and 5 of valve S.

b. Depending on the instrument model, secure the connections as follows:

• For the EASY-nLC II instrument, use a 1/4 in. open-ended wrench to tighten the fittings.

• For the EASY-nLC 1000 instrument, secure the fittings to their receiving ports as described in

“Using nanoViper Fittings” on page 83 . Take care not to overtighten

these fingertight fittings.

4. Check for leaks by running the system leak test as described in “Checking the Sample

Loop Connections for a Leak” on page 222 .

Replacing a Pressure Sensor for the PLU Pump

This topic describes how to replace a pressure sensor that is compatible with the PLU model pump (standard in EASY-nLC 1000 instruments).

Note For information about replacing a pressure sensor for a PLF model pump (standard in most EASY-nLC II instruments), see

“Replacing a Pressure Sensor for a PLF Pump” on page 117 .

Replacing the pressure sensor that is connected through solvent line tubing to a PLU model pump requires these tools and materials.

Tools

• #2 Phillips head screwdriver

• 1/4 in. open-ended wrench

• 2.5 mm L-hex wrench or ball driver

Parts and materials

• Powder-free safety gloves

• Pressure sensor, P/N LC502

CAUTION Wear powder-free gloves and safety glasses when handling parts of the LC system that come into contact with solvents.

To replace a pressure sensor that is connected to a PLU pump, follow these steps:

1.

“To remove a pressure sensor from the instrument” on page 91

2.

“To install the new pressure sensor” on page 92

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4

Routine Maintenance

Replacing a Pressure Sensor for the PLU Pump

To remove a pressure sensor from the instrument

1. Set the corresponding valve to the Center position as described in “Using the Valve

Controls” on page 204 .

2. Close down the EASY-nLC system, and then turn off the power to the instrument

(see

“Closing Down the EASY-nLC Instrument” on page 21 ).

3. Using a #2 Phillips head screwdriver, with a quarter-turn loosen the three captive screws that secure the right side panel to the instrument housing. Then remove the panel.

4. Using a 1/4 in. open-ended wrench, disconnect the stainless steel tubing attached to the inlet and outlet ports of the pressure sensor.

5. Using a 2.5 mm L-hex wrench or ball driver, remove the two screws that secure the pressure sensor L bracket to the instrument panel.

6. While holding the pressure sensor, use a 2 mm L-hex wrench to remove the two screws that secure the pressure sensor to the bracket.

7. Disconnect the pressure sensor cable from the pump.

Figure 82 shows the pressure sensor connections.

Figure 82. Pressure sensor connections

One of two screws that secure the pressure sensor to the L bracket

Screws that secure the L-bracket to the instrument panel

Solvent line connections

One of two screws that secure the pressure sensor to the L bracket

Pressure sensor cable

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Replacing a Flow Sensor

To install the new pressure sensor

1. Using a 2 mm L-hex wrench and the screws that you removed in

step 6 on page 91 ,

secure the new pressure sensor to the bracket.

2. Using a 2.5 mm L-hex wrench or ball driver and the screws that you removed in step 5

on

page 91 , reconnect the pressure sensor bracket to the interior right panel of the

instrument.

3. Reconnect the stainless steel solvent lines to the inlet and outlet ports of the pressure sensor. Using a 1/4 in. open-ended wrench, tighten the fittings.

4. Mount the right side panel to the instrument housing. Then, with a #2 Phillips head screwdriver, secure the panel by rotating the three captive screws a quarter-turn clockwise.

5. Reconnect the pressure sensor cable.

6. Run the Flush Air script as described in

“Prepare – Flush Air” on page 33

.

Replacing a Flow Sensor

The procedure for replacing the flow sensor differs between the EASY-nLC 1000 system and the EASY-nLC II system.

Depending on the instrument model, follow one of these procedures to replace a flow sensor:

“Replacing a Flow Sensor in the EASY-nLC 1000 Instrument,”

on this page

“Replacing a Flow Sensor in the EASY-nLC II Instrument” on page 96

Replacing a Flow Sensor in the EASY-nLC 1000 Instrument

Replacing a flow sensor in the EASY-nLC 1000 system requires these tools and materials.

Tools

• #2 Phillips head screwdriver

• 8 mm open-ended wrench

• 2.5 mm hex wrench or ball driver

Parts and materials

• Powder-free safety gloves

• Flow sensor A/B, P/N LC540

CAUTION Wear powder-free gloves and safety glasses when handling parts of the LC system that come into contact with solvents.

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Routine Maintenance

Replacing a Flow Sensor

To replace a flow sensor in the EASY-nLC 1000 system, follow these steps:

1.

“To remove an EASY-nLC 1000 flow sensor,”

on this page

2.

“To install an EASY-nLC 1000 flow sensor” on page 95

To remove an EASY-nLC 1000 flow sensor

1. Close down the EASY-nLC system, and then turn off the power to the instrument

(see

“Closing Down the EASY-nLC Instrument” on page 21 ).

2. Using a #2 Phillips head screwdriver, with a quarter-turn loosen the three captive screws that secure the right side panel to the instrument housing. Then remove the panel.

3. Remove the nanoViper tubing and from the inlet and outlet of the flow sensor.

Figure 83 shows the flow sensors mounted behind the bracket. The flow direction is

labeled on the bracket.

Figure 83. Flow sensors nanoViper fitting

Inline filter

Inlet end

Thermo Scientific

Outlet end

Inline filter nanoViper fitting

4. Using an 8 mm open-ended wrench, remove the inline filters from both ends of the flow

sensor (see Figure 83 ).

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Replacing a Flow Sensor

5. Remove the black cable from the flow sensor (see

Figure 84

).

Figure 84. Removing the cable from the flow sensor

6. Using an 2.5 mm hex wrench, remove the two screws that secure the flow sensor to the

bracket (see Figure 85

).

Figure 85. Removing the screws attaching the flow sensor to the bracket

Location of screws that secure flow sensor A

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7. Remove the flow sensor (see Figure 86

).

Figure 86. Removing the flow sensor

4

Routine Maintenance

Replacing a Flow Sensor

Thermo Scientific

To install an EASY-nLC 1000 flow sensor

1. Insert the new flow sensor behind the bracket.

2. Insert the two screws ( Figure 85 on page 94

) that secure the flow sensor to the bracket, and tighten the screws with a 2.5 mm hex wrench.

3. Connect the black cable to the flow sensor (see Figure 84 on page 94 ).

4. Reinstall the inline filters on both ends of the flow sensor (see Figure 83 on page 93

).

Tighten the fittings with an 8 mm open-ended wrench.

5. Reconnect the nanoViper tubing to both ends of the flow sensor (see Figure 83 on page 93 ).

6. Replace the right side panel.

7. Turn on the EASY-nLC 1000 system, and wait one hour for the flow sensor to reach the correct operating temperature.

8. Calibrate the new flow sensor by using the Flow Sensors script (see

“Calibrate – Flow

Sensors” on page 51 ).

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Replacing a Flow Sensor

Replacing a Flow Sensor in the EASY-nLC II Instrument

Replacing a flow sensor in the EASY-nLC II instrument requires these tools and materials.

Tools

• #2 Phillips head screwdriver

• 2.5 mm hex wrench or ball driver

Parts and materials

• Powder-free safety gloves

• Flow sensor A type SLG1430-150, P/N LC240

• Flow sensor B type SLG1430-025, P/N LC241

CAUTION Wear powder-free gloves and safety glasses when handling parts of the LC system that come into contact with solvents.

To replace an EASY-nLC II flow sensor, follow these steps:

1.

“To remove an EASY-nLC II flow sensor,”

on this page

2.

“To install an EASY-nLC II flow sensor” on page 99

To remove an EASY-nLC II flow sensor

1. Close down the EASY-nLC system, and then turn off the power to the instrument

(see

“Closing Down the EASY-nLC Instrument” on page 21 ).

2. Using a #2 Phillips head screwdriver, with a quarter-turn loosen the three captive screws that secure the right side panel to the instrument housing. Then remove the panel.

3. Using a 2.5 mm hex wrench, remove the two screws (see Figure 87

) that secure the flow sensor bracket to the instrument panel.

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Replacing a Flow Sensor

Figure 87. Removing the two screws that secure the flow sensor bracket to the panel

2.5 mm hex screw

4. Remove the solvent lines from both ends of the flow sensor (see Figure 88

).

Figure 88. Removing the solvent lines

Outlet ends

Thermo Scientific

Inlet ends

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Routine Maintenance

Replacing a Flow Sensor

5. Loosen the metal ring and remove the cable connected to the flow sensor (see Figure 89 ).

Figure 89. Removing the cable to the flow sensor

Cable connected to the flow sensor

6. Using a 2.5 mm hex wrench, remove the two screws that secure the flow sensor to the

bracket (see Figure 90

).

Figure 90. Removing the screws that secure the flow sensor to the bracket

Two screws per flow sensor

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Routine Maintenance

Replacing a Flow Sensor

To install an EASY-nLC II flow sensor

1. Install the new flow sensor into the bracket as follows: a. Align the new flow sensor with the mounting holes in the bracket.

IMPORTANT You must replace flow sensor A with type SLG1430-150 and flow sensor B with type SLG1430-025.

b. Insert the two screws that you removed earlier in

step 6

of

“To remove an EASY-nLC II flow sensor,”

into the bracket (see Figure 90 on page 98 ).

c.

Using a 2.5 mm L-hex wrench, tighten the two screws to secure the flow sensor to the bracket.

2. Reconnect the cable to the flow sensor, and tighten the metal ring (see

Figure 89 on page 98 ).

3. Reconnect the solvent lines to both ends of the flow sensor (see Figure 88 on page 97

).

4. Remount the bracket to the panel as follows: a. Align the bracket with the mounting holes in the panel.

b. Insert the screws that you removed from the bracket when you removed it from the panel.

c.

Using a 2.5 mm L-hex wrench, tighten the two screws to secure the flow sensor

bracket to the panel (see Figure 87 on page 97

).

5. Mount the right side panel to the instrument housing. Then, with a #2 Phillips head screwdriver, secure the panel by rotating the three captive screws a quarter-turn clockwise.

6. Turn on the EASY-nLC II instrument.

7. Wait 30 minutes for the flow sensors to reach the proper operating temperature.

8. Calibrate the new flow sensor by using the Flow Sensors script (see

“Calibrate – Flow

Sensors” on page 51 ).

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Routine Maintenance

Replacing the Hard Drive

Replacing the Hard Drive

Use this procedure to replace the hard drive in either the EASY-nLC 1000 instrument or the

EASY-nLC II instrument.

Thermo Fisher Scientific supplies several versions of the hard drive. Order the appropriate hard drive based on both the instrument model and the computer box where you are replacing the hard drive. The discontinued versions of the computer box have a PS/2 port

(see

Figure 91 ).

Figure 91. Comparison of computer ports between the obsolete and current computer boxes

Obsolete computer box Current computer box

PS/2 LAN MONITOR

USB

PS/2 port on the back panel of the obsolete computer box

LAN RS-232 MONITOR

Replacing the hard drive requires these tools and materials.

Tools Replacement part

2.5 mm hex wrench Hard drive:

Torx T-10 wrench

• EASY-nLC II instrument:

 Compatible with obsolete computer box: P/N LC281

 Compatible with current computer box: P/N LC286

–or–

• EASY-nLC 1000 instrument:

 Compatible with obsolete computer box: P/N LC581

 Compatible with current computer box: P/N LC586

To replace the hard drive, follow these steps:

1.

“To remove the hard drive” on page 101

2.

“To install the hard drive” on page 103

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Routine Maintenance

Replacing the Hard Drive

To remove the hard drive

1. Close down the EASY-nLC instrument, and then turn off the power to the instrument

(see

“Closing Down the EASY-nLC Instrument” on page 21 ).

2. Using a 2.5 mm hex wrench, loosen the two screws in the hard-drive drawer

(see

Figure 92 ).

Figure 92. Screws that secure the hard-drive drawer (obsolete computer box shown)

Two screws that secure the hard-drive drawer

3. Pull out the hard-drive drawer, as shown in

Figure 93 .

Figure 93. Pulling out the hard-drive drawer

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Replacing the Hard Drive

4. Using a Torx T-10 wrench, remove the four screws underneath the hard-drive drawer that secure the hard drive to the drawer (see

Figure 94

).

Figure 94. Removing the screws underneath the hard-drive drawer

5. Unplug the hard drive from the IDE cable (see Figure 95 ).

Figure 95. Hard drive unplugged from the IDE cable

IDE cable

The flat (IDE) cable is connected to the hard disk. The text on the hard disk is upside down, and the pins on the hard drive fit in the same place on the flat cable connector.

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Routine Maintenance

Replacing the Hard Drive

Tip If the IDE cable plug is difficult to access, remove the brass rod that holds the

cable down (see Figure 96 ). Use a Torx T-8 wrench to remove the screws. Place the

rod back in the drawer afterward.

Figure 96. Removing the brass rod that holds the cable down

Thermo Scientific

To install the hard drive

1. Insert the new hard drive.

2. Connect the hard drive to the IDE cable.

3. Insert the four screws into the holes underneath the hard-drive drawer and then, using a

Torx T-10 wrench, tighten the four screws to secure the hard drive to the drawer (see

Figure 94 on page 102

).

4. Close the hard-drive drawer.

5. Using a 2.5 mm L-hex wrench, insert and tighten the two screws that secure the drawer to the back panel (see

Figure 92 on page 101

).

6. Turn on the EASY-nLC instrument.

7. Press Vendor.

8. When prompted, enter the serial numbers for the instrument and the hard drive.

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Replacing the Hard Drive

Note When you replace the hard drive, the pumps and valves are automatically detected during the boot procedure. Because the autosampler is not detected by the boot procedure, you must add it to the Device list. After you add the autosampler to the Device list, you must calibrate the plate formats that you plan to use and the wash bottle positions, as these calibrations were stored on the old hard drive.

9. Add the autosampler to the Devices list as follows: a. Press Maintenance > Devices.

b. Press Add Device.

The Select a Device to Add dialog box opens.

c.

Locate the autosampler device and select it. d. Press Accept to add the autosampler device to the Devices list and close the Select a

Device to Add dialog box. e.

Verify that the autosampler device was added to the system by locating it in the

Devices list.

10. Calibrate the autosampler as described in Chapter 7, “Calibrating the Autosampler’s XYZ

Robot.”

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Managing the Devices List

Managing the Devices List

The Device list on the Maintenance >Devices page lists the current devices installed in the

EASY-nLC instrument.

When you add or remove a device that is not automatically detected during the boot procedure, you must add the device to or remove the device from the Devices list on the

Maintenance > Devices page. Devices that are not recognized during the boot procedure include the autosampler and the external RePlay device.

To add a device to the Device list

1. Press Maintenance > Devices.

2. Press Add Device.

The Select a Device to Add dialog box opens.

Thermo Scientific

3. Locate the device in the list and select it.

4. Press Accept to add the autosampler device to the Devices list and close the Select a

Device to Add dialog box.

5. Verify that the device was added to the system by locating it in the Devices list. Select the device in the Devices list, and review the information on the About page.

To remove a device from the Devices list

1. Press Maintenance > Devices.

2. Select the device in the Devices list.

3. Press Remove Device.

The Remove Device dialog box appears.

4. Press Accept to remove the device and return to the Maintenance > Devices page.

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Managing the Devices List

To view information about a device

1. Press Maintenance > Devices.

2. Select the device in the Devices list.

3. Press the About tab.

The About view contains information about the serial numbers and firmware versions for the rotary valves, piston (syringe) pumps, autosampler, and HPLC.

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Field Service Maintenance

This chapter provides procedures for use by Thermo Fisher Scientific field service engineers on replacing the subassemblies in the EASY-nLC instrument. Only Thermo Fisher Scientific field service engineers can perform these procedures.

CAUTION Only Thermo Fisher Scientific field service engineers can perform the procedures in this chapter.

Contents

Replacing the Built-In Computer

Replacing the Monitor

Replacing a Pressure Sensor for a PLF Pump

Replacing the ASC Autosampler

Upgrading from an ASA Autosampler to an ASC Autosampler

Replacing the Autosampler Cooler

Replacing a Rotary Valve

Replacing a Pump

Replacing a Pump PCB

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Field Service Maintenance

Replacing the Built-In Computer

Replacing the Built-In Computer

The procedure for replacing the built-in computer is the same for the EASY-nLC 1000 and

EASY-nLC II instruments, except where noted.

Replacing the computer box requires the following part and tools.

Tools

• #2 Phillips screwdriver

• Torx T-10 wrench

• Small flathead screwdriver

Parts

Computer box:

• EASY-nLC II instrument: P/N LC285

–or–

• EASY-nLC 1000 instrument: P/N LC585

To replace the built-in computer, follow these steps:

1.

“To remove the computer box,”

on this page

2.

“To install the new computer box” on page 112

To remove the computer box

1. Close down the EASY-nLC instrument (see

“Closing Down the EASY-nLC Instrument” on page 21

), and then unplug the power cable.

2. Disconnect all of the cables from the instrument’s back panel.

3. Using a #2 Phillips head screwdriver, with a quarter-turn loosen the three captive screws that secure the left side panel to the instrument housing. Then remove the panel.

4. Remove the internal cables connected to the computer box.

Note The cable connectors can be hard to loosen. You might need to wiggle the connectors carefully from side to side to loosen them.

Note Take care when you disconnect the monitor LVDS cable because it is easily damaged.

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Field Service Maintenance

Replacing the Built-In Computer

Figure 97 shows these cables in the EASY-nLC 1000 instrument and Figure 98

shows these cables in the EASY-nLC II instrument.

Figure 97. Cables connected to the computer box in the EASY-nLC 1000 instrument

LVDS cable

Figure 98. Cables connected to the computer box in the EASY-nLC II instrument

LVDS cable

Tip To remove the 12 V cables on the EASY-nLC II instrument, press the tab on the top of the plug, as shown in

Figure 99

.

Figure 99. Pressing the tab on the top of the plug of the 12 V cables

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Replacing the Built-In Computer

5. Using a Torx T-10 wrench, remove the screws that secure the back panel to the

instrument (see Figure 100

).

Note In older EASY-nLC instruments, 11 screws secured the back panel to the housing. In current EASY-nLC 1000 instruments, 3 screws secure the back panel to the housing.

Figure 100. Screws that secure the back panel to the instrument housing

USB LAN

LAN MAC ADDRESS: 00-6--E0-45-4E-28

P-BUS

IN3 OUT3 IN2 OUT2 IN1 OUT1

RS-232

HARD

DRIVE

MONITOR

I

O

110-120V▼

220-240▼

120/230 V : 50/60 Hz: 250 W

Fuse at 120 V– : T5 AL 250 V

Fuse at 230 V– : T2.5 AL 250 V

E A S Y - n L C 1 0 0 0

L C - 0 1 1 x x x

LC120

This device complies with Part 15 of the FCC Rules.

Operation is subject to the following two conditions:

(1) This device may not cause harmful interference, and

(2) This device must accept any interference received, including interference that may cause undesired operation.

This instrument is for research use only.

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Replacing the Built-In Computer

6. Using a Torx T-10 wrench, remove the four screws that secure the computer box to the

instrument (see Figure 101

).

Figure 101. Screws to remove from the computer box

7. Pull the computer box out of the EASY-nLC housing, as shown in

Figure 102 .

Figure 102. Pulling out the computer box

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Replacing the Built-In Computer

To install the new computer box

1. Insert the new computer box into the EASY-nLC housing.

2. Reconnect all of the internal cables to the computer box.

Figure 97 on page 109

shows the connections to the computer box inside the

EASY-nLC 1000 instrument.

Figure 98 on page 109

shows the connections to the computer box inside the

EASY-nLC II instrument.

3. Using a Torx T-10 wrench and the four screws that you removed in step 6 on page 111 ,

secure the computer box to the instrument (see

Figure 101 on page 111

).

4. Using a Torx T-10 wrench and the screws that you removed in

step 5 on page 110 , secure

the back panel to the instrument.

Tip Do not reconnect the left side panel to the instrument housing until after you recalibrate the autosampler.

5. Reconnect the cables to the back panel as follows:

• Reconnect the power cable to the back panel and to a laboratory power outlet.

• If the instrument was connected to a laboratory LAN port, reconnect the Ethernet cable.

• If the instrument is part of a Thermo Scientific LC/MS system, reconnect the

Ethernet cable to the back panel of the instrument, and make sure that the other end of the cable connects to the Ethernet switch for the data system hardware.

6. Turn on the EASY-nLC instrument, and log in as an administrator.

7. Add the autosampler to the Devices list (see

“Managing the Devices List” on page 105

).

8. Recalibrate the autosampler (see Chapter 7 ).

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Replacing the Monitor

Replacing the Monitor

The procedure for replacing the monitor is the same for the EASY-nLC 1000 instrument and the EASY-nLC II instrument, except where noted.

Replacing the monitor requires the following part and tools.

Tools

• Torx T-10 wrench

• 2 mm L-hex wrench or straight wrench

Part

Monitor (and adapter cable for computer boxes with a PS/2 keyboard connector as shown in

Figure 16 on page 16 ),

P/N LC282

To replace the monitor, follow these steps:

1.

“To remove the monitor,” on this page

2.

“To install the new monitor” on page 116

To remove the monitor

1. Close down the EASY-nLC instrument (see

“Closing Down the EASY-nLC Instrument” on page 21

), and unplug the power cable.

2. Depending on the instrument model, do the following:

• For the EASY-nLC II instrument, go to step 3 to remove the front panel.

• For the EASY-nLC 1000 instrument, go to step 4 to remove the top-front panel.

3. To remove the front panel from the EASY-nLC II instrument, do the following: a. Using a flat-edged tool, pry off the Thermo blue applique.

b. Using a Torx T-10 wrench, remove the 10 screws that secure the front plate to the

EASY-nLC II instrument. Then remove the plate (see Figure 103

).

c.

Go to step 5

.

Figure 103. Removing the front plate from the EASY-nLC II instrument

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Replacing the Monitor

4. To remove the top-front panel from the EASY-nLC 1000 instrument, do the following: a. Using a Torx T-10 wrench, remove the two screws that secure the top-front panel to the top back of the instrument.

b. Open the tray compartment door, and then use a Torx T-10 wrench to remove the screw that secures the top-front panel to the front of the instrument.

5. Using a 2 mm hex wrench, remove the four screws that secure the monitor to the instrument (see

Figure 104

).

CAUTION Use only hand tools with a hex tip to avoid damaging the screw heads.

Figure 104. Removing the screws from the front of the monitor

6. Using a #2 Phillips head screwdriver, with a quarter-turn loosen the three captive screws that secure the left side panel to the instrument housing. Then remove the panel.

7. Remove the USB, LVDS, and monitor cables connected to the computer box, being

careful not to damage the LVDS plug (see Figure 105

).

Figure 105. USB, LVDS, and monitor cables

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USB cable

LVDS cable

Monitor cable

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5

Field Service Maintenance

Replacing the Monitor

8. If a white plastic needle guide is installed on the back of the monitor, remove it before removing the monitor.

Figure 106

shows the white plastic needle guide on the back of the monitor.

Figure 106. White plastic needle guide

Needle guide

9. Carefully remove the monitor by pulling it straight forward (see Figure 107

).

Figure 107. Removing the monitor

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Replacing the Monitor

To install the new monitor

1. Carefully insert the new monitor into the instrument housing.

2. If you removed the white plastic needle guide on the back of the monitor, reinstall it.

Figure 106 on page 115 shows the white plastic needle guide mounted to the back of the

monitor.

3. If the computer box has a PS/2 keyboard connector on its back panel as shown in

Figure 16 on page 16 , connect the adapter cable that is supplied with the new monitor to

the monitor cable.

4. Connect the USB, LVDS, and monitor cables to the computer box, being careful not to damage the LVDS plug (see

Figure 105 on page 114

).

5. Using a 2 mm ball driver and the screws that you removed in

step 5 on page 114 , secure

the monitor to the instrument.

6. Depending on the instrument model, do the following:

• For the EASY-nLC II instrument, use a Torx T-10 wrench and the 10 screws that you removed in

step b on page 113 to secure the front plate to the housing.

• For the EASY-nLC 1000 instrument, use a Torx T-10 wrench and the three screws that you removed in

step 4 on page 114 to secure the top-front panel to the housing.

7. Mount the left side panel to the instrument housing. Then, with a #2 screwdriver, secure the panel by rotating the three captive screws a quarter-turn clockwise.

8. Plug in the power cable, and then turn on the EASY-nLC instrument.

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Replacing a Pressure Sensor for a PLF Pump

Replacing a Pressure Sensor for a PLF Pump

The procedure for replacing a pressure sensor differs between EASY-nLC 1000 instruments with PLU pumps and EASY-nLC II instruments with PLF pumps.

Note Most EASY-nLC II instruments contain the PLF pump model.

The following procedure explains how to replace a pressure sensor that is connected to a PLF pump. For instructions about replacing a pressure sensor that is connected to a PLU pump, see

“Replacing a Pressure Sensor for the PLU Pump” on page 90 .

CAUTION Wear powder-free gloves and safety glasses when handling parts of the LC system that come into contact with solvents.

Replacing the pressure sensor that is connected to a PLF pump requires these items.

Tools

• # 2 Phillips screwdriver

• 2.0 mm L-hex wrench

• 2.5 mm L-hex wrench or ball driver

• 1/4 in. open-ended wrench

Parts and materials

• Powder-free gloves

• Pressure sensor, P/N LC202

To replace a pressure sensor that is connected to a PLF pump, follow these steps:

1.

“To remove a pressure sensor that is connected to a PLF pump,” on this page

2.

“To install a pressure sensor for a PLF pump” on page 121

To remove a pressure sensor that is connected to a PLF pump

1. Set the corresponding valve to the Center position as described in “Using the Valve

Controls” on page 204 .

2. Close down the EASY-nLC II instrument (see

“Closing Down the EASY-nLC

Instrument” on page 21

), and unplug the power cable.

3. Using a #2 Phillips head screwdriver, with a quarter-turn loosen the three captive screws that secure the right side panel to the instrument housing. Then remove the panel.

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Field Service Maintenance

Replacing a Pressure Sensor for a PLF Pump

4. Disconnect the P-Bus communication cables from the pump (see

Figure 108 ).

Figure 108. P-Bus communication cables connected to the pump PCB of the PLF pump

P-Bus communication cables

5. Using a 2.5 mm L-hex wrench or ball driver, remove the four 2.5 mm hex screws that secure the PCB to the pump.

Figure 109

shows these screws. The lower right screw also connects the pressure sensor grounding cable to the PCB.

Figure 109. Four hex screws connecting the PCB to the PLF pump

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Pressure sensor grounding cable

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5

Field Service Maintenance

Replacing a Pressure Sensor for a PLF Pump

6. Gently pull the PCB toward you and rotate it to gain access to its back side

(see

Figure 110 ).

Figure 110. Turning the PCB around (PLF pump)

Pressure sensor communication cable connection to the back side of the PCB

7. Disconnect the pressure sensor communication cable, being careful not to put any stress

on the cables (see Figure 111

).

Figure 111. Disconnecting the pressure sensor communication cable (PLF pump)

Pressure sensor communication cable connection to the back side of the PCB

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Replacing a Pressure Sensor for a PLF Pump

8. While holding the pressure sensor, use a 1/4 in. open-ended wrench to disconnect the solvent lines connected to the inlet and outlet of the pressure sensor (see

Figure 112 ).

Figure 112. Disconnecting the solvent lines from the pressure sensor

Solvent lines connected to the pressure sensor

9. Using a 2.5 mm L-hex wrench or ball driver, remove the two hex screws that secure the

pressure sensor L bracket to the instrument panel (see Figure 113

).

Figure 113. Pressure sensor L-bracket screws

Screws that secure the L bracket to the instrument panel

Screws that secure the pressure sensor to the L bracket

10. Using a 2 mm L-hex wrench or ball driver, unscrew the two hex screws that secure the

pressure sensor to the L-bracket (see Figure 113

).

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Field Service Maintenance

Replacing a Pressure Sensor for a PLF Pump

To install a pressure sensor for a PLF pump

1. Disconnect and remove the pressure sensor from the EASY-nLC II instrument as

described in “To remove a pressure sensor that is connected to a PLF pump” on page 117 .

2. Using a 2 mm L-hex wrench or ball driver, insert and tighten the two hex screws to secure the pressure sensor to the L-bracket.

3. Using a 2.5 mm L-hex wrench or ball driver, insert and tighten the two hex screws

to secure the pressure sensor L-bracket to the instrument panel (see Figure 113 on page 120 ).

4. Reconnect the solvent lines to the pressure sensor (see Figure 112 on page 120 ).

5. Reconnect the pressure sensor communication cable, being careful not to put any stress

on the cables (see Figure 111 on page 119 ).

6. Gently slide in the PCB.

7. Using a 2.5 mm L-hex wrench or ball driver, insert and tighten the four 2.5 mm

hex screws to connect the PCB to the pump (see Figure 109 on page 118 ).

Note The screw in the lower right also connects the pressure sensor grounding cable to the PCB.

8. Reconnect the P-Bus communication cables to the pump (see Figure 108 on page 118

).

9. Mount the right side panel to the instrument housing. Then, with a #2 screwdriver, secure the panel by rotating the three captive screws a quarter-turn clockwise.

10. Turn on the EASY-nLC II instrument and log in as an administrator (see

“Logging In to the EASY-nLC Instrument for Maintenance Tasks” on page 20 ).

11. To draw fresh solvent into the pump, purge the pump five times by using the Purge

Solvent script on the Maintenance > Scripts page (see “Prepare – Purge Solvent” on page 32 ).

12. To remove air from the system, run the Flush Air script for the pump associated with the new pressure sensor (see

“Prepare – Flush Air” on page 33 ).

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Field Service Maintenance

Replacing the ASC Autosampler

Replacing the ASC Autosampler

The procedure for replacing the ASC autosampler is the same for the EASY-nLC 1000 instrument and the EASY-nLC II instrument. However, if your EASY-nLC instrument has an installed ASA autosampler that is irreversibly damaged, you must upgrade to an ASC autosampler, because the ASA autosampler is no longer available. For instructions on upgrading to an ASC autosampler, see

“Upgrading from an ASA Autosampler to an ASC

Autosampler” on page 128 .

CAUTION Wear powder-free gloves and safety glasses when handling parts of the LC system that come into contact with solvents.

Replacing the ASC autosampler requires these items.

Tools

• #2 Phillips screwdriver

• 3 mm L-hex wrench or ball driver

• Torx T-10 wrench

• Cutting pliers

Parts and materials

• Powder-free gloves

• 4 in. or 10 cm length tie wrap

• Autosampler (ASC), P/N LC301

To replace the autosampler, follow these steps:

1.

“To remove the ASC autosampler,” on this page

2.

“To install the ACS autosampler into an instrument that previously included this model” on page 127

To remove the ASC autosampler

1. Remove the old autosampler from the device list as follows: a. Press Maintenance > Devices.

b. Select the autosampler in the Devices list.

c.

Press Remove Device.

The Remove Device dialog box opens.

d. Press Accept.

1. Close down the EASY-nLC instrument (see

“Closing Down the EASY-nLC Instrument” on page 21

). Then unplug the power cable.

2. Using a #2 Phillips head screwdriver, with a quarter-turn loosen the three captive quarter-turn screws that secure each side panel to the instrument housing. Then remove both panels.

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Replacing the ASC Autosampler

3. Remove the computer box from the instrument as described in “To remove the computer box” on page 108 .

4. Unscrew the autosampler needle fitting, and remove or pull up the autosampler needle from the needle holder.

Figure 114

shows the autosampler needle connected to the needle holder.

Figure 114. Needle connected to the needle holder

Autosampler needle fitting

Thermo Scientific

5. Unplug the bus cables on the left side of the instrument from the autosampler cooler and horizontal XYZ-axis PCB (see

Figure 115 ).

Figure 115. Bus cables

Cooler bus cable

XYZ-axis PCB bus cable

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Replacing the ASC Autosampler

6. Cut the tie wrap attaching the cables to the vertical part of the autosampler frame, as

shown in Figure 116

.

Figure 116. Cutting the strip attaching the cables to the autosampler frame

7. Using a Torx T-10 screwdriver, remove the screws that secure the back horizontal cable bar to the autosampler frame (see

Figure 117 ).

Figure 117. Horizontal cable bar after removal of screws

Bus cable connected to valve B

Back left screw that secures the autosampler to the instrument

Horizontal cable bar

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Autosampler frame

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Field Service Maintenance

Replacing the ASC Autosampler

8. Move the cable bar to the side (this is easier to do if you also remove the bus cable to valve B). See

Figure 118 .

Figure 118. Horizontal cable bar moved aside

Valve B

Bus cable disconnected from valve B

Horizontal cable bar

Back side of the autosampler

Right side of the autosampler

9. Using a 3 mm hex wrench, remove the three screws that secure the autosampler to the bottom plate of the housing (see

Figure 119 ,

Figure 120 , and Figure 121

):

• Remove the screw from the right side of the autosampler (see

Figure 119 ).

Figure 119. Removing the screw from the right side of the autosampler

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Replacing the ASC Autosampler

• Remove the screw from the back left side of the autosampler (see Figure 120

).

Figure 120. Removing the screw from the back left of the autosampler

• Remove the screw the screw on the front of the autosampler (see

Figure 121 ).

Figure 121. Removing the front screw from the autosampler

10. Remove the autosampler from the housing through the back of the instrument.

Figure 122

shows the housing with the autosampler partially removed.

Figure 122. Housing with autosampler partially removed

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Field Service Maintenance

Replacing the ASC Autosampler

To install the ACS autosampler into an instrument that previously included this model

1. Insert the new autosampler into the housing.

2. Using a 3 mm L-hex wrench and, if necessary, a needle-nosed pliers or tweezers, insert and tighten the screws that secure the autosampler to the bottom of the housing as follows:

• Insert and tighten the screw on the back left side of the autosampler

(see

Figure 120 on page 126

).

• Insert and tighten the screw on the right side of the autosampler

(see

Figure 119 on page 125

).

• Insert and tighten the screw on the front of the autosampler

(see

Figure 121 on page 126

).

3. Insert and tighten the screws that secure the back horizontal cable bar to the autosampler frame (see

Figure 117 on page 124 ).

4. Install a new tie wrap to secure the cables to the vertical part of the autosampler frame.

5. Plug the bus cables into the autosampler cooler and horizontal XYZ-axis PCBs.

6. Reinstall the computer box as described in “To install the new computer box” on page 112 .

7. Reinsert the autosampler needle into the holder and tighten the fitting.

8. Mount the right side panel to the instrument housing. Then, with a #2 screwdriver, secure the panel by rotating the three captive screws a quarter-turn clockwise.

Tip Reconnect the left side panel to the housing after you recalibrate the autosampler.

9. Reconnect the power cable.

10. Turn on the EASY-nLC instrument.

11. Add the ASC autosampler to the Devices list on the Maintenance > Devices page as follows: a. Press Maintenance > Devices to open the Maintenance > Devices page.

b. Press Add Devices to open the Select the Device to Add dialog box.

c.

Select the ASC autosampler from the list, and then press Accept.

d. Verify that the ASC autosampler has been added to the Devices list. i.

Select the ASC autosampler in the Devices list.

ii. Check the information on the About page.

For more information about adding devices to the Devices list, see

“Managing the

Devices List” on page 105 .

12. Calibrate the autosampler as described in Chapter 7 .

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Upgrading from an ASA Autosampler to an ASC Autosampler

Upgrading from an ASA Autosampler to an ASC Autosampler

If your EASY-nLC II instrument has an ASA autosampler that is irreversibly damaged, you must upgrade to an ASC autosampler because the ASA autosampler is no longer available.

CAUTION Wear powder-free gloves and safety glasses when handling parts of the LC system that come into contact with solvents.

Upgrading the instrument by replacing an ASA autosampler with an ASC autosampler requires these items.

Tools

• 4 mm ballpoint hex wrench

• 2 mm L-hex wrench or ball driver

• 2.5 mm L-hex wrench or ball driver

• Torx T-10 wrench

• #2 Phillips screwdriver

• Tweezers

Parts and materials

• Powder-free gloves

• M5 ×16 MC hex screw

• ASC autosampler, P/N LC301

To replace the ASA autosampler with an ASC autosampler, follow these steps:

1.

“To remove the ASA autosampler from the EASY-nLC II instrument,” on this page

2.

“To install the ASC autosampler into an instrument that previously held the ASA model” on page 130

To remove the ASA autosampler from the EASY-nLC II instrument

1. Upgrade the touch-screen software to the latest version as described in “Downloading the

Latest Firmware File” on page 296

.

Note Read the release notes before you upgrade to ensure that you have the most recent version of the HPLC software.

2. Press Maintenance > Devices and remove the old autosampler from the device list.

3. Close down the EASY-nLC instrument (see

“Closing Down the EASY-nLC Instrument” on page 21

). Then unplug the power cable.

4. Remove the computer box as described in “To remove the computer box” on page 108

.

Note Removing the computer box requires the removal of the left side panel and the back panel.

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Upgrading from an ASA Autosampler to an ASC Autosampler

5. Using a #2 Phillips head screwdriver, with a quarter-turn loosen the three captive screws that secure the right side panel to the instrument housing. Then remove the panel.

6. Remove the monitor as described in “To remove the monitor” on page 113 .

7. Using a 2.5 L-hex wrench or ball driver, loosen valve W (see Figure 123

).

Figure 123. Loosened valve W

Valve W

8. Disconnect the two P-Bus communication cables from the cooler PCB and

XYZ-axis PCB on the right side of the instrument (see

Figure 129 on page 132

).

9. Remove the ASA autosampler through the side of the housing.

10. Disconnect the cables that supply power to the fans, and remove the fan assembly at the

back of the housing (see Figure 124 ).

Figure 124. Removing the fan assembly and cables

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Upgrading from an ASA Autosampler to an ASC Autosampler

To install the ASC autosampler into an instrument that previously held the ASA model

Note Because you are upgrading the instrument by adding an ASC autosampler, this upgrade procedure differs from the simple installation procedure on

page 127 .

1. As you insert the new autosampler into the housing, move the loosened valve W a little to

create the required space, as shown in Figure 125

.

Figure 125. Putting in the ASC autosampler

2. Align the holes in the autosampler plate with the three mounting holes in the bottom of the housing (see

Figure 126 ).

Figure 126. Aligning the holes in the autosampler plate with the mounting holes

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Upgrading from an ASA Autosampler to an ASC Autosampler

3. Reinstall the computer box into the EASY-nLC housing.

4. Reconnect all of the internal cables to the computer box.

Figure 98 on page 109

shows the connections to the computer box inside the

EASY-nLC II instrument.

5. Insert the P-Bus terminator between the computer box and the P-Bus communication cables as shown in

Figure 127 . It does not matter which cable the terminator connects to.

Note Do not install the connector if the computer box is labeled “Terminated,” as shown in

Figure 128 . This designation means that you do not have to install a

separate terminator cable.

Figure 127. P-Bus connector inserted between the computer box and the P-Bus communication cables

Thermo Scientific

P-Bus connector

Figure 128. Computer box with “Terminated” label

“Terminated” label

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Upgrading from an ASA Autosampler to an ASC Autosampler

6. Using a Torx T-10 wrench and the four screws that you removed in step 6 on page 111 ,

secure the computer box to the instrument (see

Figure 101 on page 111

).

7. Connect the P-Bus communication cables to the autosampler.

The longer cable connects to the cooler PCB. The shorter cable connects to the

XYZ-axis PCB. Figure 129

shows these two cables.

Figure 129. P-Bus communication cables

Figure 130

shows the boards that the P-Bus communication cables connect to.

Figure 130. Boards that the P-Bus communication cables connect to

Cooler PCB

XYZ-axis PCB

8. Using a 2.5 mm hex wrench or ball driver, mount valve W.

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Upgrading from an ASA Autosampler to an ASC Autosampler

9. Secure the autosampler to the housing as follows: a. Using a pair of tweezers, place the M5 × 16 hex screw in the back right mounting position, lifting up the autosampler in order to put the screw in place.

Figure 131 , Figure 132 , and Figure 133

demonstrate this procedure.

Figure 131. Lifting up the autosampler to position the screw

Figure 132. Placing the M5 × 16 hex screw into the back right side of the autosampler frame

Figure 133. The M5 × 16 hex screw in place

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Upgrading from an ASA Autosampler to an ASC Autosampler b. Insert the screw into the back left side of the autosampler. Use a ball driver to tighten the screw (see

Figure 134 ).

Figure 134. Securing the back left screw c.

Insert the screw into the front of the autosampler. Use a ball driver to tighten the screw (see

Figure 135 ).

Figure 135. Securing the front screw

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Upgrading from an ASA Autosampler to an ASC Autosampler

10. Place the air filter from the ASA autosampler onto the ASC cooler (see Figure 136

).

Figure 136. Placing the air filter onto the cooler

Thermo Scientific

Because the injection needles are different for the ASA and ASC autosamplers, you must install a needle arm that can be held in place underneath the mixing Tee holder.

11. Mount the aluminum needle arm under the mixing Tee holder (see Figure 137

).

Figure 137. Aluminum needle arm

Mixing Tee holder

Needle arm

12. Place the mixing Tee in the holder. Use the aluminum needle arm when you attach the needle (see

Figure 138 ).

Figure 138. Needle

Mixing Tee

Needle arm

Mixing Tee holder

13. Reinstall the monitor as described in

“To install the new monitor” on page 116 .

Tip Do not reconnect the left side panel to the instrument housing until after you recalibrate the autosampler.

14. Turn on the instrument, and then log in to the instrument as an administrator.

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Upgrading from an ASA Autosampler to an ASC Autosampler

15. Add the ASC autosampler to the Devices list on the Maintenance > Devices page as follows: a. Press Maintenance > Devices.

The Maintenance > Devices page opens.

b. Press Add Devices.

The Select the Device to Add dialog box opens.

c.

Select the ASC autosampler from the list, and then press Accept.

d. Verify that the ASC autosampler has been added to the Devices list.

For more information about adding devices to the Devices list, see

“Managing the

Devices List” on page 105 .

16. Calibrate the autosampler as described in Chapter 7, “Calibrating the Autosampler’s XYZ

Robot.”

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Replacing the Autosampler Cooler

The procedure for replacing the autosampler cooler is the same for the EASY-nLC 1000 and

EASY-nLC II instruments, but it differs between an ASC cooler and an ASA cooler.

To replace the autosampler cooler, follow the appropriate procedure:

“Replacing the ASC Cooler,”

on this page

“Replacing the ASA Cooler” on page 140

Replacing the ASC Cooler

Replacing the ASC cooler requires these items.

Tools

• Torx T-10 wrench

• 2.5 mm L-hex wrench

Parts

ASC autosampler cooling module, P/N LC310

To replace the ASC cooler, follow these steps:

1.

“To remove the ASC cooler,” on this page

2.

“To install the new ASC cooler” on page 139

To remove the ASC cooler

1. Close down and turn off the EASY-nLC instrument (see

“Closing Down the EASY-nLC

Instrument” on page 21

). Then disconnect the power cable.

2. Using a Torx T-10 wrench, remove the screws on the back cover panel, and remove the panel.

3. Access the PCB on the back of the cooler through the back of the instrument, and unplug

the P-Bus communication cables, shown in Figure 139

.

Figure 139. P-Bus communication cables

P-Bus communication cables

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Replacing the Autosampler Cooler

4. Using a 2.5 mm ball driver, remove the two 2.5 mm hex screws at the bottom of the

cooler, shown in Figure 140

.

Figure 140. Screws at the bottom of the cooler

5. Using a 2.5 mm ball driver, remove the 2.5 mm hex screw in the top center position of the cooler, as shown in

Figure 141 .

The screws do not fall out when loosened, because they are held in place by O-rings.

Figure 141. Removing the top center screw

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6. Pull out the cooler, as shown in Figure 142

.

Figure 142. Pulling out the cooler

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Replacing the Autosampler Cooler

To install the new ASC cooler

1. Insert the new cooler.

Be careful not to damage any of the cables during reassembly.

Figure 143 shows a

damaged cable.

Figure 143. Damaged cable

Damaged cable

Thermo Scientific

2. Using a 2.5 mm ball driver, insert and tighten the 2.5 mm hex screw in the top center position (see

Figure 151 on page 143

).

3. Using a 2.5 mm ball driver, insert and tighten the two 2.5 mm hex screws at the bottom of the cooler (see

Figure 150 on page 143

).

4. Plug the P-Bus communication cables into the PCB on the back of the cooler, accessing the PCB through the back of the instrument.

5. Align the back panel to the back of the instrument.

6. Using a Torx T-10 wrench, tighten the eleven screws that secure the back panel to the instrument.

7. Reconnect the power cable.

8. Turn on the EASY-nLC instrument.

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Replacing the ASA Cooler

Replacing the ASA cooler in an EASY-nLC II instrument requires these items.

Tools

• 2 mm L-hex wrench or ball driver

• 2.5 mm L-hex wrench or ball driver

• 3 mm L-hex wrench

• 5.5 mm open-ended wrench

• #2 Phillips screwdriver

Parts

ASA autosampler cooling module,

P/N EXLC252

To replace the ASA cooler, follow these steps:

1.

“To remove the ASA cooler,” on this page

2.

“To install the ASA cooler” on page 145

To remove the ASA cooler

1. Close down and turn off the EASY-nLC instrument (see

“Closing Down the EASY-nLC

Instrument” on page 21

). Then disconnect the power cable.

2. Remove the autosampler door, shown in

Figure 144 , to avoid damaging it when you

remove the hex screws underneath the adapter plate holder.

Figure 144. Autosampler door

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3. Depending on the instrument model, do one of the following:

• On the EASY-nLC II model LC100, use a 5.5 mm open-ended wrench to loosen the

nuts underneath the door (see Figure 145 ).

CAUTION Be careful removing the lid because the spring on the door can snap back.

Figure 145. Nuts beneath the lid of the EASY-nLC II instrument

• On the EASY-nLC II model LC100-2, use a 2 mm L-hex wrench to loosen the screws on the front (see

Figure 146 ).

Figure 146. Screws on the front of the EASY-nLC II instrument (model LC100-2)

Thermo Scientific

4. Using a #2 Phillips head screwdriver, with a quarter-turn loosen the three captive screws that secure the left side panel to the instrument housing. Then remove the panel.

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Replacing the Autosampler Cooler

5. Using a flathead screwdriver or your fingernail, loosen the cap that secures the ribbon cable to the controller PCB, as shown in

Figure 147 .

Figure 147. Loosening the ribbon cap on the controller PCB

Figure 148

shows a close-up view of the cap that secures the cable to the PCB.

Figure 148. Cap around the ribbon cable (close-up view)

6. Pull the ribbon cable out of the cap, as shown in Figure 149

.

Figure 149. Pulling the ribbon out of the cap

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Replacing the Autosampler Cooler

7. Using a 2.5 mm L- hex wrench, remove the lid where the ribbon cable connects to the autosampler tray (see

Figure 150 ).

Figure 150. Removing the lid

Thermo Scientific

8. Using a 2 mm hex wrench, remove the screws in the small PCB underneath the lid.

Figure 151

shows the screw in the lower right corner, and

Figure 152 shows the screw in

the middle of the left side.

Figure 151. Loosening the screw in the lower right corner

Figure 152. Loosening the screw in the middle left side

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Replacing the Autosampler Cooler

9. Using a 3 mm L-hex wrench, loosen the four hex screws underneath the adapter plate holder, and remove the holder.

Figure 153 shows the location of these screws.

Figure 153. Screws beneath the adapter plate holder

10. Lift and pull the adapter plate holder out of the autosampler, as shown in Figure 154

.

Figure 154. Pulling the adapter plate holder out of the autosampler

Note The sleeves around the holes on the bottom of the adapter plate holder can fall

out. Figure 155

shows these sleeves.

Figure 155. Hole sleeves on the bottom of the adapter plate holder

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Sleeve that has fallen out of the hole

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To install the ASA cooler

1. Align the arms of the autosampler bed, as shown in

Figure 156 .

Figure 156. Aligning the arms of the autosampler bed

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Replacing the Autosampler Cooler

2. Carefully place the new cooler in the autosampler bed, as shown in

Figure 157 .

Figure 157. Placing the new cooler in the autosampler bed

3. Verify that the cables from the fan are routed in the groove (see Figure 158

).

Figure 158. Verifying the placement of the fan cables

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Replacing the Autosampler Cooler

4. Using a 3 mm L-hex wrench, insert and tighten the four hex screws underneath the

adapter plate holder. Figure 153 on page 144 shows the location of these screws.

5. Using a 2 mm L-hex wrench, tighten the two screws in the small PCB. Figure 151 on page 143 and Figure 152 on page 143

show the locations of these screws.

6. Using a 2.5 mm L-hex wrench, mount the lid where the ribbon cable connects to the

autosampler cooler (see Figure 150 on page 143 ).

7. Insert the ribbon cable into the controller board (see

Figure 147 on page 142

and

Figure 149 on page 142 ).

8. Using a flat screwdriver or your fingernail, push the cap around the ribbon cable in place

(see

Figure 148 on page 142

).

9. Reattach the autosampler door (see Figure 144 on page 140 ).

10. Verify that the autosampler moves smoothly in and out by moving it manually.

11. Reconnect the power cable.

12. Turn on the EASY-nLC II instrument, and check the cooler temperature from the

Home > Overview page.

13. Before using the instrument, install the vial/plate adapter and perform an autosampler calibration. For instructions, see

Chapter 7, “Calibrating the Autosampler’s XYZ Robot.”

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Replacing a Rotary Valve

Replacing a Rotary Valve

The procedure for replacing a rotary valve is the same for the EASY-nLC 1000 instrument and the EASY-nLC II instrument.

CAUTION Wear powder-free gloves and safety glasses when handling parts of the LC system that come into contact with solvents.

Replacing a high-pressure valve requires these items.

Tools

• #2 Phillips screwdriver

• 1/4 in. open-ended wrench

• Torx T-10 wrench

• 2.5 mm L-hex wrench

• Flathead screwdriver

Parts and materials

• Powder-free gloves

• Switching valve:

– EASY-nLC 1000 instrument: P/N LC519

–or–

– EASY-nLC II instrument: P/N LC218

To replace a rotary valve, follow these steps:

1.

“To remove a rotary valve,”

on the page

2.

“To install a new rotary valve” on page 149

To remove a rotary valve

1. Close down the EASY-nLC instrument (see

“Closing Down the EASY-nLC Instrument” on page 21

), and then unplug the power cable.

2. Remove the computer box as described in “To remove the computer box” on page 108

.

Note Removing the computer box requires the removal of the left side panel and the back panel.

3. Using a #2 Phillips head screwdriver, with a quarter-turn loosen the three captive screws that secure the right side panel to the instrument housing. Then remove the panel.

4. From the right side of the instrument (solvent system compartment), remove the tubing from the stator on the valve that you are replacing. Be careful to avoid pulling any fused silica lines, as there is limited space around the valves.

Tip When replumbing the system, follow the solvent system schematics in

Appendix C, “Consumables and Replacement Parts.”

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Replacing a Rotary Valve

5. Using a 2.5 mm L-hex wrench, remove the four 2.5 mm hex screws that keep the valve in place inside this compartment.

As you remove the last hex screw, the valve might slip down into the autosampler compartment.

6. From the back of the instrument, disconnect the cables shown in

Figure 159

from the valve to be exchanged.

Figure 159. Cables connected to valves

7. Pull out the valve, as shown in Figure 160 .

Figure 160. Pulling out a valve

Note If there is a support collar between the body of the valve and the body of the

EASY-nLC instrument, move it to the new valve.

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Replacing a Rotary Valve

To install a new rotary valve

1. Using a small flathead screwdriver, set the rotary switch on the new valve of the PCB,

shown in Figure 161

, to the same address as the valve being replaced:

• Valve A=1

• Valve B=2

• Valve S=3

• Valve W=4

Figure 161. Valve rotary switch

Rotary switch

2. Insert the new valve, making sure that the valve PCB faces down.

3. In the back of the instrument, connect the cables shown in Figure 159 on page 148 to the

new valve.

4. Using a 2.5 mm L-hex wrench, insert and tighten the four hex screws that keep the valve in place inside the autosampler compartment.

5. In the solvent system compartment, restore the tubing and connections to the stator on the valve that was replaced.

6. Reinstall the computer box as described in step 1 through step 5 of “To install the new computer box” on page 112

.

7. Mount the left and right side panels to the instrument housing. Then, with a #2 screwdriver, secure each panel by rotating the three captive screws a quarter-turn clockwise.

8. Run the Flush Air script as described in “To run the Flush Air script” on page 34 :

• Enter 12 μL for a PLU pump (standard in the EASY-nLC 1000 instrument).

• Enter 10 μL for a PLF pump (standard in most EASY-nLC II instruments).

9. Run the Leaks script for the subsystem that you have worked on as described in

“To run the Leaks script” on page 46 .

• For valve A, select A.

• For valve B, select B.

• For valves W or S, select System.

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Replacing a Pump

Replacing a Pump

The procedure for replacing a pump differs between EASY-nLC 1000 instruments that contain PLU model pumps and EASY-nLC II instruments that contain PLF model pumps.

Note Because Thermo Fisher Scientific no longer stocks the PLF model pump as a replacement part, you must replace both pump models with the PLU model pump.

To replace a pump, follow the appropriate procedure:

“Replacing a PLU Pump,” on this page

“Replacing a PLF Pump with a PLU Pump” on page 154

CAUTION Wear powder-free gloves and safety glasses when handling parts of the LC system that come into contact with solvents.

Replacing a PLU Pump

Replacing a PLU model pump requires these tools and materials.

Tools

• #2 Phillips screwdriver

• 1/4 in. open-ended wrench

• 2.5 mm hex wrench

• 3 mm hex wrench

• Small flathead screwdriver

• Pipette for priming the pump

Parts and materials

• Powder-free gloves

• PLU model pump, P/N LC501

To replace a pump in the EASY-nLC instrument, follow these steps:

1.

“To remove a PLU model pump from an EASY-nLC instrument” on page 151

2.

“To install a PLU pump in an EASY-nLC instrument” on page 153

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To remove a PLU model pump from an EASY-nLC instrument

1. Set the valve for the pump to position 1–6 as follows: a. Press Home > Overview.

b. Press the appropriate valve icon.

c.

In the Valve dialog box, press Center.

Placing the valve in the Center position shuts off the solvent flow to the subsystem.

2. Close down and then turn off the EASY-nLC instrument.

3. Disconnect the power cable from the instrument’s back panel.

4. Using a #2 Phillips head screwdriver, with a quarter-turn loosen the three captive screws that secure the right side panel to the instrument housing. Then remove the panel.

5. Using a 1/4 in. open-ended wrench, disconnect the stainless steel tubing connected to the pump head, shown in

Figure 162 .

Figure 162. Fitting on the pump head

Fitting on the pump head

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6. Depending on the instrument model, do one of the following:

• For the EASY-nLC 1000 instrument, unplug the flow sensor cable (from pump A or B), the pressure sensor cable, and the P-Bus communication cables, shown in

Figure 163

.

Figure 163. Flow sensor cable, pressure sensor cable, and P-Bus communication cables for an EASY-nLC 1000 instrument

One of four screws that secure the pump to the panel

One of four screws that secure the pump to the panel

Flow sensor cable

Rotary switch

One of four screws that secure the pump to the panel

One of four screws that secure the pump to the panel

Pressure sensor cable

P-Bus communication cables

• For the EASY-nLC II instrument, unplug the pressure sensor cable and the

P-Bus communication cables from the pump.

The EASY-nLC II flow sensors do not have a flow sensor cable that connects to the pump.

7. Using a 3 mm ball driver or L-hex wrench, remove the four 3 mm hex screws that secure the pump to the instrument panel, and then remove the pump. Set the screws aside.

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Replacing a Pump

To install a PLU pump in an EASY-nLC instrument

1. Connect the new pump as follows: a. Using a 3 mm ball driver or L-hex wrench and the four 3 mm hex screws that you removed in

step 7 on page 152 , secure the new pump to the instrument panel.

b. Depending on the instrument model, do one of the following:

• For the EASY-nLC 1000 instrument, reconnect the flow sensor cable, the pressure sensor cable, and the P-bus communication cables.

• For the EASY-nLC II instrument, reconnect the pressure sensor cable and the

P-bus communication cables.

2. Make sure that the address setting on the pump PCB is the same as the setting for the old

PCB. Use a small flathead screwdriver to change the rotary switch address on the new pump PCB.

• Pump A =1

• Pump B = 2

• Pump S = 3

3. Reconnect the power cable to the instrument’s back panel.

4. Turn on the EASY-nLC instrument, and log in as an administrator.

5. Prime the pump as described in “Priming the Pump” on page 71

.

6. Reconnect the tubing to the pump head. Using a 1/4 in. open-ended wrench, tighten the fitting.

7. Draw fresh solvent into the solvent lines and remove air from the pump head as described in

“Removing Air After Replacing a Piston Seal or a Pump” on page 73

.

8. Depending on the pump you are replacing, do the following:

• When replacing pump A or B, run the pump leak test, and then run the flow sensor calibration (see

“Test – Leaks” on page 44 and

“Calibrate – Flow Sensors” on page 51 ).

• When replacing pump S, run the sample pickup test (see

“Test – Sample Pickup” on page 42 ).

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Replacing a Pump

Replacing a PLF Pump with a PLU Pump

You must replace an irreparably damaged PLF pump in an EASY-nLC II instrument with a

PLU pump, as Thermo Fisher Scientific no longer stocks the PLF pump as a replacement part.

When you replace a PLF pump with a PLU pump, do the following:

• Replace the pressure sensor for the pump and the high-pressure tubing associated with the pump, as the PLU pump is not compatible with the pressure sensor used in the

EASY-nLC II instrument.

• Upgrade the touch-screen software to version 3.1.4 or later, as the EASY-nLC II instrument only supports the PLU pump when running software version 3.1.4 or later.

• If you are replacing pump S in an EASY-nLC II instrument with an earlier version of the chassis, order and install the components in the Pump Mounting Kit so that the

PLU pump can fit into the pump S position.

Replacing an EASY-nLC II pump (PLF model) with a PLU pump requires these items.

Tools

• #2 Phillips screwdriver

• 1/4 in. open-ended wrench

• 2 mm hex wrench or ball driver

• 2.5 mm hex wrench or ball driver

• 13 mm open-ended wrench

• Small flathead screwdriver

• 3 mm open-ended wrench

Parts and materials

• Powder-free gloves

• Pipette

• PLU pump, P/N LC501

• Pressure sensor, P/N LC502

• Tubing between the pump and the pressure sensor:

– Pumps A and S: PEEK tubing, P/N LC212

– Pump B: prebent stainless steel tube, P/N LC215

• Tubing between the pressure sensor and the rotary valve:

– Pumps A and S: PEEK tubing, P/N LC213

– Pump B: prebent stainless steel tube, P/N LC216

To replace a PLF pump with a PLU pump in the EASY-nLC II instrument

1. Upgrade the touch-screen software to version 3.1 or later.

If the touch-screen software is older than version 2.7.8, you must first upgrade to version 2.7.8, and then upgrade to version 3.1.

For information about upgrading the software, refer to Appendix B of the EASY-nLC

Series Getting Started Guide (for the touch-screen software). You can find the touch-screen software files in the user zone on www.proxeon.com

.

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Replacing a Pump

2. Before you remove the right side panel, do the following:

a. Set the corresponding valve to the Center position as described in “Using the Valve

Controls” on page 204 .

b. Close down the EASY-nLC II instrument, and then turn off the power switch on the instrument’s back panel (see

“Closing Down the EASY-nLC Instrument” on page 21 ).

c.

Disconnect the power cable from the instrument’s back panel.

3. Using a #2 Phillips head screwdriver, with a quarter-turn loosen the three captive screws that secure the right side panel to the instrument housing. Then remove the panel.

Figure 164

shows the solvent line connections from pump A to valve A, which are made with PEEK tubing. The solvent line connections from pump B to valve B are made with stainless steel tubing.

Figure 164. Pump connections in an EASY-nLC II instrument (PEEK tubing for pump A)

Valve A

Valve A

3

2

4

1

6

5

3

2

4

1

6

5

Thermo Scientific

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Replacing a Pump

4. Disconnect the subsystem tubing between the pump head and the high-pressure valve as follows:

Note The PLF pump can have an internally or an externally threaded pump head.

The latest version of the PLF pump has an internally threaded pump head.

a. Disconnect the tubing from the pump head as follows:

• If a PEEK fitting is connected to the pump head, use a 13 mm open-ended

wrench to remove it (see Figure 165

).

Figure 165. Externally threaded pump head for a PLF model pump

PEEK fitting connected to an externally threaded pump head

• If a stainless steel fitting is connected to the pump head (see Figure 166

), use a

1/4 in. open-ended wrench to remove it.

Figure 166. Internally threaded pump head for a PLF model pump

Stainless steel fitting connected to an internally threaded pump head

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Replacing a Pump b. While holding the pressure sensor, use a 1/4 in. open-ended wrench to disconnect the tubing connected to the pressure sensor’s inlet and outlet ports (see

Figure 167 ).

Figure 167. Disconnecting the solvent lines from the pressure sensor

Solvent lines connected to the pressure sensor

Thermo Scientific c.

Using a 1/4 in. open-ended wrench, disconnect the other end of the tubing that is connected to the subsystem valve as follows:

• For valve S, disconnect the tubing from port 6.

• For valve A or B, disconnect the tubing from port 1.

5. Disconnect the pressure sensor assembly and the damaged PLF pump from the instrument panel as follows: a. Using a 2.5 mm L-hex wrench or ball driver, remove the two hex screws that secure the pressure sensor L bracket to the instrument panel (see

Figure 168 ). Then set the

two 2.5 mm hex screws aside for reuse when you reconnect the L bracket to the instrument panel. b. Using a 2 mm L-hex wrench or ball driver, unscrew the two hex screws that secure the pressure sensor to the L-bracket (see

Figure 168 ). Then set the L-bracket and the

two 2 mm hex screws aside for reuse when you mount the new pressure sensor.

Figure 168. Pressure sensor L-bracket screws

2.5 mm ball driver

Screws that secure the L bracket to the instrument panel

Screws that secure the pressure sensor to the L bracket

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Replacing a Pump c.

Disconnect the P-Bus communication cables from the pump actuator

(see

Figure 169 ).

Figure 169. Disconnected P-Bus communication cable

P-Bus communication cable d. Using a 3 mm ball driver or L- hex wrench, remove the four screws that secure the

pump to the instrument panel (see Figure 170

). e.

Set the four 3 mm screws aside for reuse in step 8a

on

page 159 .

Figure 170. Removing the four screws that secure the pump to the instrument panel

3 mm driver

6. Install the new pressure sensor as follows: a. Using a 2.0 mm L-hex wrench or ball driver and the two 2.0 mm hex screws that you set aside when you disconnected the original pressure sensor from the L-bracket, connect the new pressure sensor to the L-bracket.

b. Using a 2.5 mm L-hex wrench or ball driver and the two 2.5 mm hex screws that you set aside when you removed the L-bracket from the instrument panel, reconnect the

L-bracket to the instrument panel.

7. If you are replacing pump S in an early version of the EASY-nLC II instrument, install the

components supplied in the Pump Mounting Kit as described in “To install the bracket and adapter plate provided in the Pump Mounting Kit” on page 160 . Then continue this

installation at

step 9

.

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Replacing a Pump

8. Connect the new pump as follows: a. Using a 3 mm ball driver or L-hex wrench and the four 3 mm hex screws that you set aside when you removed the damaged pump from the instrument, secure the new pump to the instrument panel. b. Connect the pressure sensor cable from the new pressure sensor to the receptacle on the right side of the PLU pump. c.

Connect the P-Bus communication cables.

9. Check the address setting for the rotary switch. If necessary, use a small flathead screwdriver to change the rotary switch address on the new pump PCB.

• Pump A =1

• Pump B = 2

• Pump S = 3

10. Install tubing from the pump to the pressure sensor and from the pressure sensor to the valve. Use PEEK tubing for subsystems A and S. Use stainless steel tubing for subsystem B.

11. Reconnect the power cable to the instrument’s back panel, and turn on the

EASY-nLC II instrument.

12. Prime the new pump as described in

“Priming the Pump” on page 71

.

13. Remove air from the system as described in

“Removing Air After Replacing a Piston Seal or a Pump” on page 73 .

14. Make sure that both the right and left instrument panels have been installed and that the instrument has been turned on for at least 30 minutes.

15. Run the flow sensor calibration for the new pump (see

“Calibrate – Flow Sensors” on page 51 ).

16. If you replaced pump S, run the Sample Pickup script (see

“Test – Sample Pickup” on page 42 ).

The instrument is now ready for use.

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Replacing a Pump

To install the bracket and adapter plate provided in the Pump Mounting Kit

Note Some earlier versions of the EASY-nLC II instrument cannot accommodate a

PLU pump in the pump S position without modification.

To provide space for the pump outlet tubing in these instruments, you must install the new flow sensor bracket and the pump adapter plate supplied in the Pump

Mounting Kit. The new flow sensor bracket places the flow sensors in a higher position and the pump adapter plate places the pump in a lower position than their original positions on the instrument panel. This repositioning makes space for the tubing that is connected to the pump head.

1. If you have not already done so, follow step 1 on page 154

through

step 6

on

page 158 of

the previous procedure, “To replace a PLF pump with a PLU pump in the EASY-nLC II instrument,”

to upgrade the touch-screen software, gain access to the solvent system compartment, remove the damaged pump, and replace the pressure sensor.

2. Disconnect the tubing from the inlet and outlet ports of the flow sensors.

3. Replace the flow sensor bracket with the new flow sensor bracket supplied in the Pump

Mounting Kit as follows: a. Using a 2.5 mm hex wrench, remove the two screws that secure the flow sensor bracket to the instrument panel (see

Figure 171 ).

b. Set the two 2.5 mm screws aside for reuse when you secure the new flow sensor bracket.

Figure 171. Removing the two screws that secure the flow sensor bracket to the panel

2.5 mm hex wrench c.

Using a 2.0 mm hex wrench or ball driver, remove the four screws that secure the two

flow sensors to the flow sensor bracket (see Figure 172

). d. Set the 2.0 mm hex screws aside for reuse when you secure the flow sensors to the new flow sensor bracket.

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Replacing a Pump

Figure 172

shows the four 2.0 mm hex screws that secure the flow sensors to the flow sensor bracket and the original flow sensor bracket.

Figure 173 shows the flow sensors

removed from the flow sensor bracket. The flow sensor cables are still connected to the instrument.

Figure 172. Screws that secure the flow sensors to the bracket

Figure 173. Flow sensors removed from the flow sensor bracket

Two screws per flow sensor

Thermo Scientific e.

Using a 2.5 mm hex wrench and the screws that you set aside when you removed the flow sensors from the original flow sensor bracket, connect the flow sensors to the new flow sensor bracket supplied in the Pump Mounting Kit.

Note The mounting holes in the new flow sensor bracket are lower than the mounting holes in the original flow sensor bracket. When you connect the new bracket to the instrument panel, it is positioned higher than the original bracket, allowing more space for the tubing that is connected to the pump head.

Figure 174. New flow sensor bracket (with lower mounting holes)

Lower mounting holes in the new flow sensor bracket

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Replacing a Pump

4. Using a 2.5 mm hex wrench and the screws that you set aside when you removed the original flow sensor bracket from the instrument panel, connect the new flow sensor bracket to the instrument panel.

5. Reconnect the tubing to the inlet and outlet ports of the flow sensors. Be careful when installing the fused silica line to flow sensor B, as the end of the tubing is fragile.

CAUTION Take care when you connect the fused-silica line to the outlet port of flow sensor B. The end of the tubing is fragile and easily damaged.

6. Using a 2.5 mm L-hex wrench or ball driver and the four screws provided in the Pump

Mounting Kit, secure the pump adapter plate to the instrument panel.

Figure 175. Securing the pump adapter plate to the instrument panel

Pump adapter plate

7. Install the PLU pump in the pump S position as follows: a. Using a 3 mm ball driver or L-hex wrench and the four 3 mm hex screws that you set aside when you removed the damaged pump from the instrument panel, secure the new pump to the pump adapter plate.

b. Connect the pressure sensor cable from the new pressure sensor to the receptacle on the right side of the PLU pump. c.

Connect the P-bus communication cables.

8. To complete the pump installation, start from

step 9 on page 159 of the previous

procedure,

“To replace a PLF pump with a PLU pump in the EASY-nLC II instrument.”

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Replacing a Pump PCB

Replacing a Pump PCB

The procedure for replacing the pump PCB differs between the PLU model pump and the

PLF model pump.

Note The PLU pump is standard in the EASY-nLC 1000 instrument. The PLF pump is standard in most EASY-nLC II instruments.

Figure 8 on page 9 shows these two pump

models.

To replace a pump PCB, follow the appropriate procedure:

“Replacing a Pump PCB in the PLU Pump,” on this page

“Replacing a Pump PCB in the PLF Pump” on page 167

Replacing a Pump PCB in the PLU Pump

Replacing the pump PCB on the PLU model pump requires these items.

Tools

• #2 Phillips screwdriver

• 2.5 mm hex wrench

• Small flathead screwdriver

Parts

Pump PCB, P/N LC574

To replace the pump PCB on the PLU pump, follow these steps:

1.

“To remove a pump PCB from the PLU model pump,”

on this page

2.

“To replace a pump PCB on the PLU model pump” on page 166

To remove a pump PCB from the PLU model pump

1. Close down the EASY-nLC 1000 instrument as described in

“Closing Down the

EASY-nLC Instrument” on page 21 .

2. Disconnect the power cable from the instrument’s back panel.

3. Using a #2 Phillips head screwdriver, with a quarter-turn loosen the three captive screws that secure the right side panel to the instrument housing. Then remove the panel.

4. Disconnect the P-Bus communication cables, the flow sensor cable, and the pressure sensor cable from the pump.

Figure 176 on page 164 shows these cables connected to the pump.

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Replacing a Pump PCB

5. Using a 2.5 mm L-hex wrench or ball driver, loosen the four captive screws on the PCB shield (see

Figure 176 ). Then remove the PCB shield (see Figure 177

).

Figure 176. Four PCB shield screws (PLU model pump)

Flow sensor cable

Pressure sensor cable

P-Bus cables

The four captive screws remain in place (see Figure 177

).

Figure 177. Removing the PCB shield (PLU model pump)

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Replacing a Pump PCB

6. Slide the PCB forward and away from the four mounting studs.

7. Disconnect the motor cable on the back of the pump PCB (see

Figure 178 ).

Figure 178. Motor cable connected to the pump PCB (PLU model pump)

Motor cable

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Replacing a Pump PCB

To replace a pump PCB on the PLU model pump

1. Using a small flathead screwdriver, reset the address on the rotary switch on the pump

PCB (see Figure 179

) as follows:

• Pump A =1

• Pump B = 2

• Pump S = 3

Figure 179. PCB rotary switch

PCB rotary switch

2. Plug the motor cable into the back of the PCB (see Figure 178 on page 165 ).

3. Insert the new pump PCB onto the pump.

4. Install the PCB shield.

5. Using a 2.5 mm L-hex wrench or ball driver, tighten the four 2.5 mm hex screws in the

PCB shield.

6. Reconnect the P-Bus communication cables, the flow sensor cable, and the pressure

sensor cable (see Figure 176 on page 164

) to the pump.

7. Mount the right side panel to the instrument housing. Then, with a #2 screwdriver, secure the panel by rotating the three captive screws a quarter-turn clockwise.

8. Reconnect the power cable to the back panel of the instrument.

9. Turn on the instrument.

10. Wait for 30 minutes to allow the temperature in the housing to stabilize.

11. Calibrate the flow sensor by following the automated script described in “Calibrate –

Flow Sensors” on page 51 .

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Replacing a Pump PCB

Replacing a Pump PCB in the PLF Pump

Replacing a pump PCB (PLF model pump) in the EASY-nLC II instrument requires these items.

Tools

• #2 Phillips screwdriver

• 2.5 mm hex wrench

• Small flathead screwdriver

Parts

Pump PCB, P/N LC274

To replace the PCB on a PLF pump, follow these steps:

1.

To remove the pump PCB from the PLF model pump (EASY-nLC II instrument)

2.

To install the pump PCB on a PLF pump (EASY-nLC II instrument)

To remove the pump PCB from the PLF model pump (EASY-nLC II instrument)

1. Close down the EASY-nLC II instrument as described in “Closing Down the EASY-nLC

Instrument” on page 21

.

2. Disconnect the power cable from the instrument’s back panel.

3. Using a #2 Phillips head screwdriver, with a quarter-turn loosen the three captive screws that secure the right side panel to the instrument housing. Then remove the panel.

4. Disconnect the P-Bus communication cables (see Figure 169 on page 158 ) from the

pump actuator.

5. Unscrew and remove the four 2.5 mm hex screws that connect the PCB to the pump.

6. Disconnect the pressure sensor grounding cable (yellow wire).

7. Gently slide the PCB outward and turn it around to gain access to its back side.

8. Disconnect the pressure sensor communication cable. Take care to avoid putting any stress on the cables.

9. Disconnect the motor cable. Again, take care to avoid putting any stress on the cables.

10. Remove the PCB.

11. Go to the next procedure

“To install the pump PCB on a PLF pump (EASY-nLC II instrument).”

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Replacing a Pump PCB

To install the pump PCB on a PLF pump (EASY-nLC II instrument)

1. Remove the damaged PCB as described in the previous procedure, “To remove the pump

PCB from the PLF model pump (EASY-nLC II instrument)” on page 167 .

2. Using a small flathead screwdriver, reset the address on the rotary switch on the new pump PCB as follows:

• Pump A =1

• Pump B=2

• Pump S=3

3. Reconnect the motor cable to the back of the PCB (see

Figure 178 on page 165

).

4. Mount the new PCB onto the pump.

5. Secure the PCB to the pump as follows:

• Slide the four screws into the holes in the four corners of the PCB.

• Slide the connector at the end of the pressure sensor grounding cable under the screw in the lower-right corner of the PCB.

• Using a 2.5 mm hex wrench, tighten the four screws.

Figure 180. PLF pump with the pressure sensor cable connected

9

0 1

2

P-Bus cable

Pressure sensor grounding cable

P-Bus cable

Motor cable routed to the back of the PCB

CAUTION To avoid damaging the PCB, make sure that the grounding connector does not touch any other components.

6. Reconnect the P-Bus communication cables.

7. Calibrate the flow sensor by following the automated script described in “Calibrate –

Flow Sensors” on page 51 .

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6

Troubleshooting

To troubleshoot problems with the EASY-nLC instrument, follow the troubleshooting tips and procedures in this chapter.

Contents

Troubleshooting Tips

Using the Direct Controls for Troubleshooting and Maintenance

Troubleshooting a Pump that Fails the Flush Air Script

Troubleshooting a Pump that Fails the Leaks Script

Running a System Leak Test

Troubleshooting the Results of the System Leak Test

Identifying a Leaking Check Valve

Troubleshooting a System Blockage

Troubleshooting the Autosampler Aspiration and Calibration

Checking Sample Pickup

Troubleshooting Communication Problems

Verifying that the LC/MS System Is Properly Grounded

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Troubleshooting

Troubleshooting Tips

Troubleshooting Tips

To troubleshoot problems with the EASY-nLC instrument, see these topics:

“Autosampler Problems” on page 171

“Contact Closure Problems” on page 172

“Instrument Startup Problems” on page 172

“Delayed Elution” on page 173

“Excessive Duration or Higher Pressure for the Column Equilibration and Sample

Loading Steps” on page 178

“System Reaches Its Maximum Pressure During the Gradient” on page 180

“Sample Signal Weak or Absent” on page 183

“Slow or No Pressure Increase in Subsystem A or B” on page 187

“Errors Reported by the Xcalibur Data System” on page 189

“Carryover” on page 194

“Spray Issues” on page 196

“Chromatographic Performance” on page 197

“Device Failures” on page 198

“Miscellaneous” on page 202

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Troubleshooting Tips

Autosampler Problems

Table 9 lists troubleshooting tips for the autosampler.

Note For information about troubleshooting sample pickup, see

“Troubleshooting the

Autosampler Aspiration and Calibration” on page 261

.

Table 9. Autosampler troubleshooting tips

Symptom

The needle cannot penetrate the plastic film on the microtiter plate.

Possible causes

The needle tip is damaged or bent.

Action

Do the following:

1. Check the software configuration for the adapter tray.

2. Eject the tray.

XYZ robot error

3. Remove the left side panel and visually inspect the needle tip.

4. Do one of the following:

• If the needle tip is bent or damaged, replace it

(see

“Replacing the Autosampler Needle” on page 84 ).

Possible step loss on one of the motors because the movement of the autosampler’s

XYZ arm is blocked

The error repeats without any visible reason.

• If there is any residue on the needle tip, wipe the needle tip with a lint-free tissue soaked in

LC/MS-grade ethanol.

Remove the obstacle and try again. You might need to calibrate the autosampler for more precise penetration of the microtiter plate or the wash bottles.

For information about calibrating the autosampler, see

Chapter 7, “Calibrating the Autosampler’s XYZ Robot.”

Contact Thermo Fisher Scientific:

[email protected]

A server warning message about the plate format for the autosampler configuration pops up when you submit a batch.

The plate format has been deleted from the system configuration. For information about deleting a plate format, refer to the EASY-nLC Series

Getting Started Guide.

[email protected]

If you want to run the batch, you must create the plate format again.

For information about selecting, creating, and deleting plate formats, see

“Managing Plate Formats” on page 274

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Troubleshooting

Troubleshooting Tips

Contact Closure Problems

Table 10

contains troubleshooting tips for contact closure problems.

Table 10. Contact closure troubleshooting tips

Symptom

The mass spectrometer is waiting for a contact closure signal.

Possible causes

Incorrect contact closure setup

Action

If you are running two-way contact closure (with feedback from the mass spectrometer), set the contact closure Protocol setting to Two-way on the

Configuration > Connections page.

The EASY-nLC instrument is waiting for a ready signal from the mass spectrometer.

If the mass spectrometer is waiting for a signal, set the contact closure State setting to Open.

If you are running one-way contact closure (that is, the EASY-nLC instrument sends a start signal to the mass spectrometer, but feedback from the mass spectrometer is ignored), set the contact closure

Protocol setting to One-way on the Configuration >

Connections page.

Instrument Startup Problems

Table 11

contains troubleshooting tips for instrument startup problems.

Table 11. Startup troubleshooting tips

Symptom

Dark monitor

Possible causes

Broken or open main power fuse

Action

1. Turn on the power to the instrument.

Defective monitor

2. Remove the right side panel of the EASY-nLC instrument.

3. If there is no light on the pumps, turn off the EASY-nLC instrument, replace the right side panel, and check the fuse.

Attach an external monitor to the VGA connector (labeled

MONITOR) on the back panel of the instrument (see Figure 16 on page 16

).

Startup bar is no longer proceeding.

Damaged instrument Turn off the EASY-nLC instrument, wait 5 seconds before turning it on again, and then wait approximately 10 minutes for the restart process to finish. In some cases this recovers the system.

Contact Thermo Fisher Scientific if the problem recurs:

[email protected]

[email protected]

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Troubleshooting

Troubleshooting Tips

Delayed Elution

The most common cause of delayed elution is a worn rotor seal in valve B, causing valve B to leak.

Table 12

lists the common causes of delayed elution. These topics show the effect of leaks and swept volume on the pressure and flow traces:

“Leaks that Cause Delayed Elution” on page 174

“Introduced Swept Volume” on page 176

Table 12. Common causes of delayed elution

Symptom

Delayed elution

Possible cause Action

Leak in the solvent system 1. Run the Leaks script for the system (see

“Test – Leaks” on page 44 ).

Incorrect tubing installed

2. Locate the leak by following “Troubleshooting the Results of the System Leak Test” on page 221

.

1. Remove the right side panel of the instrument.

2. Verify that the tubing matches the solvent system schematic.

• For the EASY-nLC II instrument, see page 314 .

• For the EASY-nLC 1000 instrument, see

page 316 .

3. Replace the incorrect tubing with the specified tubing.

Check the tubing connections. Swept volume introduced by incorrect tubing connections

In the EASY-nLC II instrument, you can inadvertently introduce swept volume (extra volume in the solvent path) by not seating the tubing against the bottom of the receiving port.

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Troubleshooting

Troubleshooting Tips

Leaks that Cause Delayed Elution

When there is a leak in the system upstream from flow sensor B, part of the solvent stream from pump A exits the mixing Tee toward flow sensor B, pushes solvent B back toward valve B, and partly fills the solvent line between the mixing Tee and flow sensor B during the column equilibration and sample loading steps (see

Figure 181 ).

When the gradient step begins, instead of pushing solvent B into the mixing Tee, pump B initially pushes this small volume of solvent A, delaying the start of the actual gradient.

Figure 181. Negative flow of solvent B toward valve B during the column equilibration and sample loading steps

Pressure sensor

Positive flow

Mixing Tee

Pump B

2

3

1

6

4 5

Valve A

Flow sensor A

Pressure sensor

Negative flow

Pump B

2

3

1

6

4 5

Valve B

Flow sensor B

A leak in valve B or the solvent line between valve B and flow sensor B can allow solvent A to enter the solvent path between the mixing

Tee and valve B during the column equilibration and sample loading steps.

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Troubleshooting Tips

Figure 182

shows the flow and pressure traces for pump B for a system with a leak upstream of flow sensor B. The measured flow rate is negative rather than 0 nL/min during the column equilibration and sample loading steps.

Figure 182. Flow trace for pump B when there is a leak upstream of flow sensor B

Negative flow rate during the column equilibration and sample loading steps

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Troubleshooting

Troubleshooting Tips

Introduced Swept Volume

Normally when solvent B (acetonitrile with 0.1% formic acid) reaches the column, the back pressure measured by pressure sensor A drops to less than one-half the amount measured for solvent A (water with 0.1% formic acid) alone. If the back pressure does not drop by more than 50 percent when solvent B reaches the column, the solvent system might contain additional swept volume introduced by poor tubing connections.

The additional swept volume increases the gradient delay volume so that the gradient reaches the column later.

Figure 183

shows the pressure dip that occurs when the gradient starts to reach the column.

Figure 184

shows the pressure dip for the sample chromatographic method, but occurring two minutes later due to the added gradient delay volume.

Figure 183. Normal pressure dip for pressure sensor A as the gradient reaches the column

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EASY-nLC Series Troubleshooting and Maintenance Guide

The pressure starts to drop at approximately

25 minutes as 100% solvent B reaches the column.

Thermo Scientific

6

Troubleshooting

Troubleshooting Tips

Figure 184. Pressure dip begins later when the system has introduced swept volume

Thermo Scientific

The pressure starts to drop at approximately

27 minutes as

100% solvent B reaches the column.

Back pressure is proportional to the viscosity of the solvent mixture. If you are not running a water/acetonitrile gradient, the pressure profile of your gradient run will differ.

Table 31

on

page 311 lists the viscosities for two-solvent mobile phases consisting of a

water/methanol mixture or a water/acetonitrile mixture.

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Troubleshooting

Troubleshooting Tips

Excessive Duration or Higher Pressure for the Column Equilibration and Sample

Loading Steps

Table 13

lists possible causes for the following:

• When the column equilibration and sample loading steps are controlled by a specified pressure, the duration of these steps is longer than expected.

• When these steps are controlled by a specified flow, the pressure is higher than expected during the column equilibration step, the sample loading step, or both steps.

During the sample loading and column equilibration steps, pump A delivers the user-specified volume of solvent A to the system at the user-specified flow rate or maximum pressure or the maximum system pressure. When the columns or the Column Out line is clogged, the system must lower the flow rate to maintain the pressure at the maximum pressure specified in the method. The lower flow rate increases the column equilibration time, the sample loading time, or both.

Table 13. Common causes of a longer than expected column equilibration time or sample loading time

Symptom

When the method specifies a set flow rate for the column equilibration, sample loading, or both of these steps, the pressure is higher than usual.

Possible cause

Clogged column

When the method specifies a maximum pressure for one or both of these steps, the duration of these steps is longer.

Clogged system

Action

Run the Back Pressure script for pump A

(see

“Test – Back Pressure” on page 48 ).

If the test passes, one of the columns is probably clogged and must be replaced.

Run the Back Pressure script for pump A.

If the test fails, go to

System Blockage” on

“Troubleshooting a page 249 .

Figure 185

and

Figure 186 show the flow rate and pressure traces for pump A for a normal

run and a run with a partially clogged column, respectively. The flow rate for a run with a clogged column is lower than the flow rate for a normal run. The lower flow rate increases the duration of the column equilibration and sample loading steps.

Note The specifications for the precolumns used to acquire the data in

Figure 185

and

Figure 186 are as follows:

• 2 cm length

• 100 μm ID

• 5 μm particle size

• C18

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Figure 185. Normal run

Sample loading step (10 μL/min)

Precolumn equilibration step (10 μL/min)

Figure 186. Run with a partially blocked column

6

Troubleshooting

Troubleshooting Tips

Thermo Scientific

Sample loading step (4 μL/min)

Precolumn equilibration step (3.5 μL/min)

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Troubleshooting

Troubleshooting Tips

System Reaches Its Maximum Pressure During the Gradient

If the pressure trace shows maximum pressure during the gradient, review the possible causes in

Table 14

.

Table 14. Possible causes of maximum pressure spikes during a run

Symptom

The system reaches its maximum pressure during a run.

Possible cause

Clogged column

Action

Run the Back Pressure script for pumps A and B.

Clogged system

If the test passes, one of the columns is probably clogged and needs to be replaced.

Run the Back Pressure script for pumps A and B.

If the test fails, go to

“Troubleshooting a System

Blockage” on page 249 .

Excessively high flow rate Reduce the flow rate in the method.

When a column or the Column Out line is clogged, the back pressure can increase to the maximum system pressure for pump A, pump B, or pumps A and B during the gradient

(see

Figure 187 ).

Figure 187. Back pressure shown reaching the maximum system pressure for pumps A and B

Maximum pressure reached for pump A

Maximum pressure reached for pump B

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Troubleshooting

Troubleshooting Tips

When only the flow path between pump B and the mixing Tee has a blockage, the back pressure on pump B increases and differs from the back pressure on pump A.

Figure 188

shows the typical profile for a blockage in the solvent system between pump B and the mixing Tee.

Figure 188. Back pressure shown reaching the maximum pressure for pump B only

Maximum pressure reached for pump B

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Troubleshooting

Troubleshooting Tips

Figure 189

shows the flow path between pump B and the mixing Tee.

Figure 189. Solvent lines between pump B and the mixing Tee

1

Valve B

2

Mixing Tee

Pressure sensor

3

Pump B

Connected to check valve assembly B

2

3

1

6

4 5

Flow sensor B

4

No.

EASY-nLC II instrument

1

2

3

Stainless steel tubing and fittings

Stainless steel tubing and fittings

PEEKsil tubing

4

• Stainless steel fitting connected to port 6 of valve B

• PEEK fitting connected to the flow sensor inlet

Fused-silica solvent line

• PEEK fitting connected to the flow sensor outlet

• PEEK fitting connected to the mixing Tee

EASY-nLC 1000 instrument

Stainless steel tubing and fittings

Stainless steel tubing and fittings nanoViper nanoViper

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Troubleshooting

Troubleshooting Tips

Sample Signal Weak or Absent

If the sample signal is missing or weak, review the troubleshooting tips in

Table 15 .

Table 15. Possible causes of a weak or absent signal

Symptom

The sample signal is missing or weak.

Possible cause

Sample not aspirated

(See

Figure 190 on page 184 .)

Damaged or incorrect column

(See Figure 191 on page 185 .)

Action

Run the Sample Pickup script. If the Sample

Pickup script fails, see “Troubleshooting the

Autosampler Aspiration and Calibration” on page 261 .

Make sure that you have installed the correct column or columns.

Check the back pressure on the columns by running the column equilibration scripts.

Run the Leaks test on the system.

Leak in the system

(See Figure 192 on page 186 .)

If the test fails, see “Troubleshooting the Results of the System Leak Test” on page 221

.

Incorrect column configuration Check the column configuration.

1. Press Maintenance > Devices.

2. In the Devices list, select EASY-nLC

(HPLC).

3. Press the Properties tab.

4. Clear the One Column Setup check box for a two-column setup or select this check box for a one-column setup.

For more information, refer to the EASY-nLC

Series Getting Started Guide.

Incorrect solvent in W3 bottle Make sure that the W3 bottle contains solvent A.

Incorrect sample loop configuration

Check the loop configuration on the Properties page for the EASY-nLC HPLC device on the

Maintenance > Devices page.

Make sure that the configured loop volume matches the installed sample loop.

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Weak or Absent Signal—Sample Not Aspirated

A pressure drop at the start of the sample loading step indicates that pump S has introduced

air into the sample loop (see Figure 190

).

When the method uses pressure to control the flow rate, the pressure should return to the specified level within approximately 4 seconds after the sample is loaded onto the column. If the pressure does not return to the specified level within approximately 4 seconds, pump S has probably introduced air into the system.

Figure 190. Typical pump A profile for a system with air in the sample loop

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Pressure drop at the start of the

Sample Loading step

Thermo Scientific

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Weak or Absent Signal—Defective Precolumn

When compared to the Pump A graph for a normal run shown in Figure 185 on page 179

,

Figure 191

shows the reduced time for the precolumn equilibration and sample loading steps when the precolumn is damaged and has lost a significant portion of its packing material.

The flow rate for the column equilibration and sample loading steps is higher (>15 μL/min) and the duration is shorter because the precolumn is exerting less back pressure.

Note The specifications for the precolumns used to acquire the data in

Figure 185

and

Figure 186 are as follows:

• 2 cm length

• 100 μm ID

• 5 μm particle size

• C18

Figure 191. Typical pump A profile for a damaged precolumn

The measured flow rate is > 15 μL/min.

Thermo Scientific

Column equilibration step

Sample loading step

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Weak or Absent Signal—Leak Upstream of Flow Sensor B

Figure 192

shows a typical pump B profile for a system with a leak upstream of flow sensor B.

Flow sensor B displays a negative flow during the column equilibration and sample loading steps.

If the gradient is short, a leak upstream of flow sensor B will cause a complete lack of signal because solvent B is not reaching the columns within the duration of the gradient.

Figure 192. Typical pump B profile for a leak upstream of flow sensor B

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Negative flow during the column equilibration and sample loading steps

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Troubleshooting Tips

Slow or No Pressure Increase in Subsystem A or B

A lack of pressure or a slow increase in pressure in subsystem A or B is often caused by a worn piston seal, a worn rotor seal, or a leaking tubing connection.

Note A subsystem includes the pump, the solvent line that connects the pump to the pressure transducer, the solvent line that connects the pressure transducer to the valve, and the valve.

Table 16. Troubleshooting low back pressure in subsystems A or B

Symptom Possible cause Action

Back pressure is low in subsystem A or B.

Air in the pump Run the Purge Solvent script for the affected pump with 2 iterations.

Then, run the Flush Air script. If the Flush Air script fails, see

“Troubleshooting a Pump that Fails the Flush Air Script” on page 212 .

Leak in the pump Run the Leaks script for the affected pump. If the Leaks script fails, see

“Troubleshooting a Pump that Fails the Leaks Script” on page 216 .

Figure 193

shows the leak test profile for pump A. Even when the flow rate is set to the maximum allowable value, the pressure for pump A does not increase.

Figure 193. Leak test profile for pump A

Thermo Scientific

Extremely low pressure

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Troubleshooting Tips

When subsystem A has a large leak, pump A can run out of solvent during a sample run, resulting in the server error message shown in

Figure 194

.

Figure 194. Server error message displayed when pump A runs out of solvent

When subsystem B has a large leak or is filled with air, the gradient fails to start because the pressure cannot be increased to reach the starting conditions (see

Figure 195

).

Figure 195. Subsystem B leak

Pressure remains low

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Troubleshooting Tips

When pump B cannot generate sufficient pressure to start the gradient, the server error

message shown in Figure 196

appears.

Figure 196. Server error message displayed when pump B cannot generate sufficient pressure to start the gradient

Errors Reported by the Xcalibur Data System

Table 17

lists the possible causes of the common errors reported by the Xcalibur data system.

Table 17. Possible causes of errors reported by the Xcalibur data system (Sheet 1 of 2)

Symptom Possible cause

An Acquisition Server failure prevents the sequence from running

(see

Figure 197 on page 190

).

Lost connection between the

EASY-nLC instrument and the external data system computer

Action

Check the Ethernet connections between the

EASY-nLC instrument and the data system computer.

Exit the Xcalibur data system. Then, open the Thermo EASY-nLC Configuration dialog box and click Test Connection.

The Acquisition Server displays the message that the EASY-nLC device has reported a failure during the

Prepare For Run step.

Injection volume is greater than the loop size minus 2 μL

If necessary, restart the EASY-nLC system.

Check the loop configuration on the

Properties page for the EASY-nLC HPLC device on the Maintenance > Devices page.

(See Figure 198 on page 191 .)

Make sure that the configured loop volume matches the installed sample loop.

Lowercase letter used in the sequence table for vial position

Change the letters to capitals.

Unable to submit samples. The following error message appears:

Invalid Autosampler Vial Position

(See Figure 199 on page 191 .)

Incorrect plate configuration

(See Figure 200 on page 192 .)

Select the correct autosampler plate in the

Thermo EASY-nLC Configuration dialog box.

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Table 17. Possible causes of errors reported by the Xcalibur data system (Sheet 2 of 2)

Symptom

The mass spectrometer remains in the Waiting for Contact Closure state and data acquisition does not start (see

Figure 201 on page 193

).

Possible cause

Contact closure configured incorrectly

Failing contact closure relay due to incorrect grounding

Action

Press Configuration > Connections.

Select the correct MS configuration and settings.

Check accordingly to “Verifying that the

LC/MS System Is Properly Grounded” on page 268 .

Figure 197

shows the Acquisition Server message that appears when the EASY-nLC instrument loses communication with the data system computer during a sequence run. The sequence pauses until you intervene.

Figure 197. Lost connection between the EASY-nLC instrument and the Acquisition Server

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Troubleshooting Tips

Figure 198

shows the Acquisition Server message that appears during the Prepare for Run step when the requested injection volume is greater than the configured loop size minus 2 μL.

Figure 198. Incorrect injection volume

A requested injection volume of

20 μL is too large for a 20 μL loop.

Figure 199

shows the Acquisition Server message that appears when you incorrectly use a lowercase letter to define the vial or well position. For the EASY-nLC device, you must use capital letters in the alphanumeric description of the vial or well position.

Figure 199. Incorrect vial position nomenclature

Incorrect use of a lowercase letter

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Troubleshooting Tips

Figure 200

shows the Home Page error message that appears when the sequence list contains a well plate position and the autosampler configuration is set up for the vial adapter plate format.

Figure 200. Incorrect vial position for the vial adapter plate format

Invalid vial position

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Troubleshooting Tips

Figure 201

shows that the mass spectrometer is waiting for a start signal from the EASY-nLC device before it starts data acquisition. The mass spectrometer remains in the Waiting for

Contact Closure state until it receives the start signal. The EASY-nLC instrument sends the start signal when the run gradient step begins.

Figure 201. Waiting for Contact Closure error

Thermo Scientific

Mass spectrometer state

EASY-nLC method step

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Carryover

Table 18

lists the possible causes of carryover.

Table 18. Possible causes of carryover

Symptom

Carryover from one run to the next

Possible cause

Incomplete elution of peptides from column

Action

Add extra time to the end of the gradient or add a high-organic hold to the end of the gradient.

Insufficient washing of needle and loop

Dead volumes in the column setup

To determine if the high-organic hold is long enough, make sure that you see a pressure drop during the hold period (see

Figure 202 and

Figure 203 ).

Add extra wash steps to the method. Consider using an organic solvent for one of the custom wash solvents.

Remake the connections in the column setup.

Figure 202

shows a gradient run with a high-organic hold at the end of the run that is too short for the column to see the high-organic solvent, as displayed by the lack of a pressure drop at the end of the run.

Figure 202. Gradient run without an adequate high-organic hold at the end of the run

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Troubleshooting Tips

Figure 203

shows the effect of a high-organic hold at the end of the gradient run on pressure.

The pressure continues to drop until the column equilibrates to the high-organic content.

Figure 203. Gradient run with an adequate high-organic hold at the end of the run

The gradient reaches

100% solvent B.

Thermo Scientific

The pressure begins to drop when the high-organic mobile phase reaches the column.

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Spray Issues

Table 19

lists the possible causes of an unstable spray.

Table 19. Possible causes of an unstable spray

Symptom

Unstable spray

Possible cause Action

Post-column outgassing

Poor connection in either the liquid junction or the

DirectJunction

Degas the solvents.

Check all of the connections.

Incorrect MS settings or emitter position

Dirty emitter

Optimize the MS tune file for the desired flow rate and the emitter position.

Clean the emitter with an organic solvent such as methanol, acetonitrile, or ethanol.

Use only LC/MS-grade solvents.

Poor high-voltage connection Check all of the high-voltage connections.

Reconnect as necessary.

Air movement from an air conditioning vent

Deflect the air from the vent, or use a shield to protect the ion source.

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Troubleshooting Tips

Chromatographic Performance

Table 20

lists troubleshooting tips for common chromatography problems.

Table 20. Troubleshooting tips for chromatographic performance

Symptom

Broad peaks

Possible cause

Dead volume between the column and emitter

Action

Check the connections.

Damaged column

Replace the fittings if needed.

Inspect the column, especially the outlet.

Poor retention time reproducibility Temperature fluctuations in the laboratory

Replace the column if it is damaged.

Monitor the laboratory temperature.

Stabilize the temperature by keeping the laboratory doors closed and by ensuring that the air conditioning system is kept constant at all times.

Back flow on flow sensor B

Spikes seen in flow sensor B during valve shifts

Do not run the EASY-nLC instrument with the panels removed.

Run the Leaks script for Pump A, Pump B, and System.

Replace defective piston seals, rotor seals, and solvent lines as indicated.

Use the scripts to check for a leak in the B subsystem (Leak test Pumps A + B, and

System).

Replace defective seals, rotors, and solvent lines as needed.

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Device Failures

Table 21

lists troubleshooting tips for devices failures.

Table 21. Troubleshooting tips for device failures

Symptom

The pump icon for a pump that is not functioning turns a pink color on the Home >

Overview page.

Possible cause Action

Communication failure to the pump.

Could be the pump or the PC BUS cable.

Check that the cables are connected correctly.

See Figure 204 on page 199 .

The valve icon for a valve that is not functioning turns a pink color on the Home >

Overview page.

Communication failure to the valve.

Could be the valve or the PC BUS cable

Check that all of the cables are connected correctly.

See Figure 205 on page 200 .

The autosampler icon turns a pink color on the Home >

Overview page.

See Figure 206 on page 201 .

Communication failure on the cooling unit.

Could be the PC box, a damaged cable, or a loose connection.

Check that all of the cables are connected correctly.

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A pump icon turns pink when the pump loses communication with the EASY-nLC computer.

The pumps are serially connected, so if pump B loses communication, all of the pumps lose communication. Pump B is the first pump in the series.

Communication between the pumps and the embedded computer can be lost when the cables are loose, when the embedded computer fails, or when the pump PCB fails.

Figure 204

shows the pink pump icons on the Home > Overview page.

Figure 204. Pink pump icons on the Home > Overview page

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A pink valve icon means that the communication to the unit has been lost. Be aware that the valves are serially connected, so a failing valve might cause other valve icons to turn pink as well.

Communication between the valves and the embedded computer can be lost when the cables are loose, when the embedded computer fails, or when the valve PCB fails.

Figure 205

shows the pink valve icons on the Home > Overview page.

Figure 205. Pink valve icons on the Home > Overview page

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An autosampler icon turning pink means that the communication to the unit has been lost.

Figure 206

shows the pink autosampler icon on the Home > Overview page.

Figure 206. Pink autosampler icon on the Home > Overview page

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Miscellaneous

For additional issues that might arise while using the EASY-nLC instrument, consult these miscellaneous troubleshooting tips (

Table 22 ).

Table 22. Miscellaneous troubleshooting tips (Sheet 1 of 2)

Symptom

Copying log files to a USB memory stick failed.

Possible causes Action

Memory stick in wrong format Format the memory stick in FAT/FAT16 format.

Memory stick not recognized by the instrument

Use one of the memory sticks supplied by

Thermo Fisher Scientific, or try plugging the stick into the other USB port.

Copying to the USB memory stick does not stop.

Too much data to be copied

Contact Thermo Fisher Scientific if neither action works: us.customer-support.analyze

@thermofisher.com

eu.techsupport.cms

@thermofisher.com

Remove the memory stick from the EASY-nLC instrument and press Ignore when the error message appears.

Forgot the admin password.

System is running slower and slower.

Contact Thermo Fisher Scientific for an instrument check-up: us.customer-support.analyze

@thermofisher.com

eu.techsupport.cms

@thermofisher.com

If you are already logged in to the EASY-nLC instrument, Thermo Fisher Scientific can extract the password from the system in two ways:

• If the network is set up for remote access, press Maintenance > Support and press

Connect.

Memory filled with graph data from long gradient run

System overload caused by other reasons

–or–

• Run a factory reset.

Press Home > Graph and disable the graphs by pressing No Graph in each graph window.

Restart the application or power down/power up from Maintenance > Scheduling.

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Table 22. Miscellaneous troubleshooting tips (Sheet 2 of 2)

Symptom

Valve – unknown position

Possible causes

The valve’ s duty cycle requires calibration.

Action

Run the Valve Check script (see “Test – Valve

Check” on page 47

).

Unhandled error or SVG

(scalable vector graphics) error

Program error in the software release

This script is only valid for EASY-nLC II instrument valves with a serial number less than

V-009999.

Contact your local Thermo Fisher Scientific field service engineer if the reported overshoot is greater than 6.

In most situations you can continue your work by pressing Ignore or OK. More serious error situations might require restarting the HPLC.

To improve on the software quality, e-mail details to: us.customer-support.analyze

@thermofisher.com

eu.techsupport.cms

@thermofisher.com

Or, connect to the remote support server.

See

“Connecting the EASY-nLC Instrument to the

Support Server” on page 288 .

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Using the Direct Controls for Troubleshooting and Maintenance

Using the Direct Controls for Troubleshooting and Maintenance

You can access the direct controls for the instrument’s hardware components by pressing the component icons on the Home > Overview page.

These topics describe how to use the direct controls:

Using the Valve Controls

Using the Pump Controls

Using the Autosampler Controls

Using the Valve Controls

You can change the positions of the instrument’s three-position valves from the

Home > Overview page. For information about the valve positions, see “Six-Port Rotary

Valves” on page 12 .

To change the position of a valve

1. On the Home > Overview page, press the valve icon for the valve that you want to control.

The Valve dialog box opens (see

Figure 207

). The readback box lists the current valve position.

Figure 207. Valve dialog box

Readback box

2. Press the button for the position that you want the valve to switch to: 1–2, Center, or 1–6.

The readback box displays the new position.

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Using the Pump Controls

You can refill and empty the pumps from the Home > Overview page.

IMPORTANT Use the pump’s direct controls for troubleshooting.

To avoid contaminating the pumps, take care when you use these controls:

• For pump A or B, make sure that its corresponding valve is in position 1–2 before you refill the pump.

• For pump S, before you empty or fill the pump, use the autosampler’s direct controls to insert the autosampler needle into the W4 or W3 bottle, respectively.

Tip To prepare the EASY-nLC instrument for operation, use the Prepare – Purge Solvent

script to flush the solvent lines (see “Prepare – Purge Solvent” on page 32

). The Purge

Solvent script places the valves in the appropriate positions for filling and emptying pumps A and B and places the autosampler needle in the appropriate wash and waste bottles for filling and emptying pump S.

To work with the pump’s direct controls, follow these procedures:

Opening the Direct Controls for the Pumps

Using the Direct Controls To Fill or Empty Pump A or B

Using the Direct Controls To Fill or Empty Pump S

Opening the Direct Controls for the Pumps

Each pump is represented by a pump icon on the Home > Overview page.

To open the pump dialog box

On the Home > Overview page, press the pump icon for the pump that you want to control.

Its dialog box opens (see

Figure 208 ). The Position readback box displays the current

pump position from 0 μL (empty) to 140 μL (full).

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Figure 208. Pump dialog box

Volume box

Flow rate box

Readback boxes

Note Because pump S does not have an associated flow sensor, the Pump S dialog box does not include a Real Flow readback box.

Using the Direct Controls To Fill or Empty Pump A or B

Before using the direct controls to fill or empty pump A or B, make sure that the valves are in the appropriate position. For information about controlling the position of the valves, see

“Using the Valve Controls” on page 204 .

IMPORTANT Before you fill pump A or B, make sure that the pump’s associated valve is in position 1–2. You can contaminate the pump by attempting to fill the pump when its associated valve is in position 1–6.

To fill pump A or B with solvent

1. On the Home > Overview page, check the position of the valve that controls the solvent flow direction for the pump of interest.

2. Make sure that the valve is set to position 1–2.

When valve A or B is in the 1–2 position, the corresponding pump draws solvent from

the solvent bottle on top of the instrument (see Figure 209 ).

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Figure 209. Position of the rotary valve when filling a pump

Check valve assembly

Solvent bottle A

2

3

1

A

4

6

5

Waste beaker

Pump A

3. Press the pump icon for the pump you want to control.

The Pump dialog box opens (see Figure 208

).

4. In the flow rate box, enter the filling flow rate: –0.01 to –300 μL/min.

For best results, enter a flow rate of –100 μL/min to fill the pump.

5. In the volume box, enter the volume that you want pump A or B to draw.

6. Press Start.

The pump draws solvent until it draws the requested volume or reaches the full position, whichever occurs first.

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Using the Direct Controls for Troubleshooting and Maintenance

To empty pump A or B to waste

1. Check the position of the pump valve on the Home > Overview page.

2. Do one of the following:

• To empty pump A or B directly to waste, make sure that its valve is in position 1–2.

• To empty pump A or B to waste by way of valve W, place the pump valve in position 1–6 and valve W in position 1–6.

Valve

A or B

S

W

Position

1–2

1–6

1–2

1–6

1–2

1–6

Solvent flow

Empties to the waste beaker.

Passes through valve S and into the Column Out line.

Bypasses the sample loop.

Passes through the sample loop.

Passes through the installed columns.

Exits the Column Out line to waste for a one column setup or passes through the precolumn and then exits the system to waste for a two-column setup.

3. Press the pump icon for the pump you want to control.

The Pump dialog box opens (see Figure 208 on page 206

).

4. In the flow rate box, enter the flow rate: 0.01 to 300 μL/min.

When valve A or B is in position 1–6, the maximum system pressure limit constrains the maximum flow rate.

5. In the volume box, enter the volume that you want the pump to empty to waste.

6. Press Start.

The pump empties until it pumps the requested volume or reaches the empty position, whichever occurs first.

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Using the Direct Controls for Troubleshooting and Maintenance

Using the Direct Controls To Fill or Empty Pump S

Before using the direct controls to fill or empty pump S, make sure that the autosampler needle is inserted into the appropriate wash or waste bottle.

IMPORTANT To avoid contaminating pump S, take care to place the autosampler needle in the appropriate wash bottle before filling the pump.

To fill pump S

1. Make sure that a wash bottle with the appropriate solvent is installed in position W3.

2. Send the autosampler needle to the appropriate position as follows: a. On the Home > Overview page, press the autosampler icon.

The Autosampler dialog box opens (see

Figure 210 on page 210 ).

b. Press W3.

The autosampler needle moves to the W3 position.

c.

Press Needle Down.

The autosampler needle descends into the W3 bottle.

d. Press Close to close the Autosampler dialog box.

3. Draw solvent into pump S as follows: a. Press the pump S icon.

The Pump S dialog box opens. b. In the flow rate box, enter the filling flow rate: –0.01 to –300 μL/min.

For best results, enter a flow rate of –40 μL/min to fill pump S.

c.

In the volume box, enter the volume that you want pump S to draw.

d. Press Start.

The pump draws solvent until it draws the requested volume or reaches the full position, whichever occurs first.

To empty pump S to waste

1. Make sure that a wash bottle with a needle wash insert is installed in position W4.

2. Send the autosampler needle to the appropriate position as follows: a. On the Home > Overview page, press the autosampler icon.

The Autosampler dialog box opens (see

Figure 210 on page 210 ).

b. Press W4.

The autosampler needle moves to the W4 position.

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Using the Direct Controls for Troubleshooting and Maintenance c.

Press Needle Down.

The autosampler needle descends into the W4 bottle.

d. Press Close to close the Autosampler dialog box.

3. Dispense solvent from pump S as follows: a. Press the pump S icon.

The Pump S dialog box opens. b. In the flow rate box, enter the flow rate: 0.01 to 300 μL/min. c.

In the volume box, enter the volume that you want the pump to empty to waste. d. Press Start.

The pump empties until it pumps the requested volume or reaches the empty position, whichever occurs first.

Using the Autosampler Controls

You can control the XYZ robot and the cooler temperature from the Home > Overview page.

To open the Autosampler dialog box

Press the autosampler icon on the Home > Overview page.

The Autosampler dialog box opens (see

Figure 210 ).

Figure 210. Autosampler dialog box

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Vial or well position box

Temperature input box

Temperature readback box

Thermo Scientific

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Using the Direct Controls for Troubleshooting and Maintenance

To start the calibration procedure for the current plate format

1. Press Goto Calibration.

The Tools view for the autosampler on the Maintenance > Devices page opens.

2. To calibrate the XYZ robot for the plate format, follow the instructions in “Calibrating

Plates” on page 280

.

To move the needle to a vial or well position

1. In the Vial or Well box, enter a vial or well position.

2. Press Goto.

To move the needle to a wash bottle position

Press W1, W2, W3, or W4.

To change the cooler temperature

1. Enter a temperature in the temperature input box.

2. Press Set.

3. Monitor the temperature change in the temperature readback box.

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Troubleshooting a Pump that Fails the Flush Air Script

Troubleshooting a Pump that Fails the Flush Air Script

If pump A, B, or S fails the Flush Air script and the Output page displays one of the following messages, a leak or blockage could be preventing solvent from entering the pump.

• Aborting unable to build pressure

• Aborting check solvent level

Note For information about the Flush Air script, see

“Prepare – Flush Air” on page 33 .

The following procedure systematically checks for a blockage in the check valve assembly, in the solvent line that connects the pressure sensor to the valve, and in the pump head.

CAUTION Wear powder-free gloves and safety glasses when handling parts of the LC system that come into contact with solvents.

To determine why the pump has failed the Flush Air script

1. If you have not already done so, remove the right side panel of the instrument.

2. Depending on the pump, do one of the following:

• For pumps A and B, go to

step 3

.

• For pump S, go to step 4 on page 213

.

3. For pumps A and B, determine whether the check valve assembly is functioning as follows: a. Run the Purge script for the pump that has failed (see

“Prepare – Purge Solvent” on page 32 ).

b. Check that solvent is being aspirated and dispensed correctly through the Teflon™ lines.

• If the pump is drawing solvent during the fill portion of the purge cycle, the solvent-side check valve is working properly.

• If the pump is dispensing solvent into the waste beaker during the empty portion of the purge cycle, the waste-side check valve is working properly.

Note If a check valve is not working properly, replace it as described in

“Replacing the Check Valves” on page 80 .

c.

If the check valve assembly is functioning properly, go to step 4 .

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Troubleshooting a Pump that Fails the Flush Air Script

4. Check whether the pump can draw solvent through the tubing that connects the pressure sensor to the rotary valve as follows: a. Using a 1/4 in. open-ended wrench, disconnect the solvent line from the rotary valve

(see

Figure 211 ).

• For pumps A or B, disconnect the solvent line from port 1 of the valve.

• For pump S, disconnect the solvent line from port 6 of the valve.

Figure 211. Rotary valve layout for the EASY-nLC 1000 instrument

Port 6 of valve S

Port 1 of valve A

Port 1 of valve B

Thermo Scientific b. Place the free end of the solvent line into a small vial that contains the appropriate solvent. Use solvent A for pump A and solvent B for pump B.

c.

Run the Purge Solvent script and check that solvent is being aspirated from the vial into the pump.

For information about the Purge Solvent script, see “Prepare – Purge Solvent” on page 32 .

d. Depending on whether the pump aspirated the solvent, do one of the following:

• If the pump does not aspirate solvent from the vial, go to

step 5

.

• If the pump aspirates solvent from the vial, the problem is probably resolved. To

verify that the system is working, go to step 4e .

e.

Reconnect all of the solvent lines, and then run the Flush Air script again.

f.

Depending on the results of the Flush Air script, do one of the following:

• If the Flush Air script fails, go to

step 6 on page 214

.

• If the Flush Air script passes, you have completed this troubleshooting procedure.

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Troubleshooting a Pump that Fails the Flush Air Script

5. Check whether the tubing that connects the pressure sensor to the valve is blocked as follows: a. Reconnect the solvent line to port 1 of the valve. b. Using a 1/4 in. open-ended wrench, disconnect the fitting connected to the top of the pressure sensor.

c.

Pipette solvent into the port on the top of the pressure sensor.

CAUTION Be careful not to spill solvent on the PCBs.

• For the PLU pump in either the EASY-nLC 1000 or EASY-nLC II instrument, the LED panel cover for each pump protects the PCB from accidental contact when the instrument’s right side panel is removed.

However, the cover does not form a waterproof seal with the pump body so that the PCBs are exposed to solvent leaks from above.

• For the PLF pump in the EASY-nLC II instrument, the PCBs are completely exposed.

d. Run the Purge Solvent script and check that the pump is aspirating solvent through the pressure sensor. e.

Depending on whether solvent is aspirated, do one of the following:

• If the pump does not aspirate solvent, go to step 6 .

• If the pump aspirates solvent, the tubing that connected the pressure sensor to the valve is damaged. Do the following: i.

Using a 1/4 in. open-ended wrench, disconnect this tubing from the valve, and install a new solvent line.

ii. To confirm that installing new tubing fixes the problem, run two iterations of the Purge Solvent script. Then, run the Flush Air script with a threshold of 10 μL for the PLF pump or 12 μL for the PLU pump.

6. To determine if the problem is in the pump, do the following: a. Reconnect the solvent line to the top of the pressure sensor and tighten the fitting.

b. Disconnect the solvent line from the pump head by doing one of the following:

• If a PEEK fitting is connected to the pump head, use a 13 mm open-ended wrench, to disconnect it.

• If a stainless steel fitting is connected to the pump head, use a 1/4 in. open-ended wrench to disconnect it.

c.

Pipette the appropriate solvent into the port in the pump head (see

Figure 65 on page 71 ).

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Troubleshooting a Pump that Fails the Flush Air Script d. Run the Purge script and check that the pump is aspirating solvent from the pump head into the pump.

• If the pump does not aspirate solvent from the pump head, the problem is in the pump.

• If the pump does aspirate solvent from the pump head, replace the tubing that connects the pump to its pressure sensor.

7. If the problem is in the pump, verify that the piston is not dirty, broken, or scratched and replace the piston seal as described in

“Maintaining the Syringe Pumps” on page 60

.

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Troubleshooting a Pump that Fails the Leaks Script

Troubleshooting a Pump that Fails the Leaks Script

Follow this procedure to determine why the pump failed the Leaks script.

Note For information about running the Leaks script see

“Test – Leaks” on page 44

.

In the Value column on the Parameters page of the Leaks script, select A to test pump A or B to test pump B.

This procedure systematically checks whether the following problems caused the Leaks script failure:

• Excess air in the system

• Tubing connections between the pump and the pressure sensor

• Tubing connections between pressure sensor and the valve

• A worn piston seal (in the pump head)

• A worn rotor seal (in the subsystem valve)

CAUTION Wear powder-free gloves and safety glasses when handling parts of the LC system that come into contact with solvents.

To troubleshoot a pump that has failed the Leaks script

1. Run the Flush Air script for the failed pump (see “Prepare – Flush Air” on page 33 ).

Enter the following value for the Flush Volume parameter:

• For the PLF model pump, enter 10 μL.

• For the PLU model pump, enter 12 μL.

2. Do one of the following:

• If the Output page reports one of these messages, go to

“Troubleshooting a Pump that Fails the Flush Air Script” on page 212 .

– Aborting unable to build pressure

– Aborting check solvent level

• If the script passes, go to step 3 .

3. Run the Leaks script for the failed pump or both pumps (see “Test – Leaks” on page 44

).

• If the Leaks script fails, the leak is either in the pump, the valve, or the connecting lines or fittings. Go to

step 4 .

• If the Leaks script passes, excess air in the system was the source of the problem.

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4. To determine whether the leak is between the pressure sensor and the valve or the pressure sensor and the pump, do the following: a. Remove the fitting from the top of the relevant pressure sensor, and replace it with a blanking nut. Tighten the nut with a 1/4 in. open-ended wrench.

• EASY-nLC II blanking nut: LC223

• EASY-nLC 1000 blanking nut: LC523 b. Repeat the Test – Leaks script.

c.

Depending on the result of the Test – Leaks script, do one of the following:

• If the Test – Leaks script passes, go to step 5 , as the piston seal is leak tight and

the leak is between the pressure sensor and the valve.

• If the Test – Leaks script fails, go to

step 6

, as the leak is between the pressure sensor and the pump.

5. To locate a leak between the pressure sensor and the valve, do the following: a. Remove the blanking nut and replace the original fitting.

b. Manually check the fittings between the pressure sensor and the valve, and confirm that they are leak tight.

If the fittings are leak tight, the valve might be the source of the leak.

c.

If the valve is leaking, replace the worn rotor seal and clean or replace the stator in the affected valve (see

“Maintaining the Rotary Valves” on page 74 ).

d. Run the Test – Leaks script for the relevant pump and confirm that you have fixed the leak.

6. To locate a leak between the pressure sensor and the pump, do the following: a. Remove the blanking nut and replace the original fitting.

b. Confirm that the fittings between pump and the pressure sensor are leak tight. c.

Repeat the Test – Leaks script. d. If the script fails again, the cause might be a worn piston seal.

e.

If you suspect that the leak is caused by a worn piston seal, go to “Maintaining the

Syringe Pumps” on page 60 .

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Running a System Leak Test

Running a System Leak Test

To identify and locate a leak downstream from valves A and B, run the system leak test

(Test – Leaks script with System selected), and then take appropriate action as described in

Table 24 on page 221 .

Table 23

lists the materials required to identify and locate leaks. Because this troubleshooting procedure requires you to handle the solvent lines, you must wear gloves and safety glasses.

CAUTION Wear powder-free gloves and safety glasses when handling parts of the LC system that come into contact with solvents.

Table 23. Materials required for running the system leak test and troubleshooting leaks

Instrument

EASY-nLC II

Union

P/N SC600

Plug or blind fitting

EASY-nLC 1000

Zero-dead-volume union for

1/32 in. OD tubing

P/N SC900 Thermo Scientific Dionex™

P/N 6040.2303

HPLC zero-dead-volume union for

1/16 in. OD tubing

To run a system leak test, follow these steps:

1.

“To prepare the instrument for a system leak test” on page 219

2.

“To run a system leak test” on page 219

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Running a System Leak Test

To prepare the instrument for a system leak test

1. Run the Flush Air script for pump A as follows: a. Press Maintenance > Scripts. b. In the Category list, select Prepare. In the Name list, select Flush Air. c.

Press the Parameters tab.

d. In the Flush Threshold [μL] box, enter the following value:

• For the PLF model pump (EASY-nLC II instrument), enter 10.

• For the PLU model pump (EASY-nLC 1000 instrument), enter 12.

Figure 25 on page 33

shows the parameters for the Flush Air script.

e.

Press Start.

The script starts and the Output page opens.

2. Depending on the results of the Flush Air script displayed on the Output page, do the following:

• If the script passes, run a system leak test as described in

“To run a system leak test” on page 219

.

• If one of these messages appears, go to

“Troubleshooting a Pump that Fails the Flush

Air Script” on page 212 .

– Aborting unable to build pressure

– Aborting check solvent level

CAUTION Wear powder-free gloves and safety glasses when handling parts of the LC system that come into contact with solvents.

To run a system leak test

1. If you have not already done so, run the Flush Air script to prepare the instrument for a system leak test (see

“To prepare the instrument for a system leak test” on page 219 ).

2. Connect the Column Out and Waste In lines with the appropriate union (see Table 23

on

page 218 ).

For the EASY-nLC 1000 instrument, connect these lines to the union as follows: a. Insert the nanoViper fittings on the ends of the Column Out and Waste In lines into the stainless steel union, and then slowly tighten the fittings until you feel resistance.

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Running a System Leak Test b. Tighten both fittings by an additional 45 degrees (1/8-turn). Do not tighten the fittings by more than a 1/4-turn (see

“Using nanoViper Fittings” on page 83

).

CAUTION Because the fittings seal against each other inside the union, do not tighten them by more than 90 degrees (1/4-turn). Overtightening the fittings can irreparably damage their sealing surfaces.

Figure 212

shows the connection between the Column Out and Waste In lines for the

EASY-nLC 1000 instrument.

Figure 212. Column Out line connected to the Waste In line (EASY-nLC 1000 instrument) nanoViper fitting

Union

(P/N SC900)

IMPORTANT Follow the instructions in

“Using nanoViper Fittings” on page 83

to ensure that the nanoViper fittings are securely connected to the union.

3. Open the Test – Leaks script as follows: a. Press Maintenance > Scripts. b. In the Category list, select Test. c.

In the Name list, select Leaks.

4. Set the parameters for the system leak test as follows: a. Press the Parameters tab.

b. In the Value list, select System.

5. Press Start.

The script starts and the Output page opens. During the duration of this script, the

EASY–nLC instrument pressurizes the flow paths highlighted with a dotted red line in

Figure 35 on page 45

, and it monitors the flow sensors and pump piston movements to identify and localize leaks. When the script ends, the Output page displays a message.

6. Depending on the message displayed on the Output page of the Test – Leaks script, take the appropriate action as listed in

Table 24 .

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Troubleshooting the Results of the System Leak Test

Table 24

lists the actions required to troubleshoot the results of a system leak test.

Tip The

“Using nanoViper Fittings Quick Reference Guide” on page 323 describes how

to return the solvent lines in the flow path of the EASY-nLC 1000 instrument to atmospheric pressure. Print this reference guide to refer to while troubleshooting leaks coming from the nanoViper tubing connections.

Table 24. Output messages for a system leak test and actions to take when this test fails

Message

System tight.

(Sensor A <100nl/min)

Flow sensor A should be calibrated.

The 0 nL/min point is off by more than ±30 nL/min, so the software cannot determine leaks.

Flow sensor B should be calibrated.

The 0 nL/min point is off by more than ±30 nL/min, so the software cannot determine leaks.

Check valve S for leaks. (Flow sensor A >100 nL/min)

Check the sample loop for leaks.

Action

No action is required; however, the system leak test does not test for a a leak in valve B or pump B.

Rerun the system leak test. If the same message appears, run the Flow Sensor script to calibrate flow sensor A (see

“Calibrate – Flow Sensors” on page 51 ).

Rerun the system leak test. If the same message appears, run the Flow Sensor script to calibrate flow sensor B (see

Go to

“Calibrate – Flow Sensors”

“Checking Valve S for a Leak” on

on page 51

page 222 .

).

Go to “Checking the Sample Loop Connections for a

Leak” on page 222 .

Go to “Locating a Leak in the Solvent Path from

Valve B to Flow Sensor B” on page 223 .

Check the B solvent line (valve to flow sensor) and valve B for leaks.

(Flow sensor B ±30 nL/min)

Check from pump A to flow sensor A for leaks. (Pump A flow loss >1000 nL/min)

Check from valve S to valve W, including valve W, for leaks.

Check solvent lines between flow sensor A and valve S, including the mixing Tee and flow sensor B, for leaks.

Go to

Pump A to Flow Sensor A”

Go to

Go to

“Locating a Leak in the Solvent Path from

“Locating a Leak In or Between Valve S and

Valve W” on page 226 .

“Locating a Leak Between the Flow Sensors and Valve S” on page 234

.

on page 224 .

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Checking Valve S for a Leak

The system leak test returned the following message (see Table 24 on page 221 ):

Check valve S for leaks. (Flow sensor A >100 nL/min)

The most likely cause of this test result is a leak in the flow path between ports 3 and 4 of valve S.

To fix the leaking valve

1. Clean the rotor seal and stator in valve S with methanol and a lint-free swab

(see

“Cleaning the Rotor Seal and Stator” on page 74

).

2. Repeat the system leak test script and if it fails again, replace the rotor seal in valve S

(see

“Replacing the Rotor Seal” on page 77 ).

3. To return the EASY-nLC instrument to normal operation as part of an LC/MS system, secure the right side panel, reconnect the Column Out line to the column assembly, and then reconnect the Waste In line to the venting Tee.

Checking the Sample Loop Connections for a Leak

The system leak test returned the following message (see Table 24

):

Check the sample loop for leaks.

The sample loop connections are leaking, or the rotor seal in valve S is worn.

To fix the leaking sample loop connections or the leaking valve

1. Depending on the instrument model, do one of the following:

• For an EASY-nLC II instrument, use a 1/4 in. open-ended wrench to tighten the sample loop fittings connected to ports 2 and 5 on valve S.

• For an EASY-nLC 1000 instrument, return the instrument to atmospheric pressure as

described in the “Using nanoViper Fittings Quick Reference Guide” on page 323 .

Then individually disconnect and reconnect the nanoViper fittings connected to ports 2 and 5 on valve S. Take care not to overtighten these fingertight fittings.

2. Rerun the system leak test (see

“Running a System Leak Test” on page 218 ).

3. If the script still fails, clean the rotor seal and stator using methanol and a lint-free swab.

Then rerun the system leak test as described in “Cleaning the Rotor Seal and Stator” on page 74 ).

4. If the problem persists, replace the rotor seal (see

“Replacing the Rotor Seal” on page 77 ).

5. If the problem persists after replacing the rotor seal, replace the sample loop

(see

“Replacing the Sample Loop” on page 89 ).

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Locating a Leak in the Solvent Path from Valve B to Flow Sensor B

The system leak test returned the following message (see Table 24 on page 221 ):

Check the B solvent line (valve B to flow sensor B) and valve B for leaks.

Flow sensor B has detected solvent flow toward valve B. This negative flow rate is caused by a leak in valve B, or in the connections to port 6 of valve B and the flow sensor inlet.

Note When the system is leak tight and valve B is in the Center position, the solvent flow through flow sensor B from pump A should be negligible. A negative flow rate of up to –30 nL/min is within the normal range. A higher negative flow rate indicates a leak that could affect the retention time of analytes.

Figure 213

shows a reading for flow sensor B of –60 nL/min. This means there is a leak in the solvent path from valve B to flow sensor B.

Figure 213. Solvent line between valve B and flow sensor B

Possible leaking connections

Valve B

–60 nL/min

2

3

1

6

4 5

Flow sensor B

To troubleshoot a leak in the solvent line connections between valve B and flow sensor B

1. Depending on the instrument model, do one of the following:

• For the EASY-nLC II instrument, do the following:

– Using a 1/4 in. open-ended wrench, tighten the stainless steel fitting connected to port 6 of valve B.

– Hand tighten the PEEK fitting connected to the flow sensor inlet.

• For the EASY-nLC 1000 instrument, return the solvent system upstream of valve S to atmospheric pressure as described in the

“Using nanoViper Fittings Quick Reference

Guide” on page 323 . Then individually disconnect and reconnect the nanoViper

fittings connected to port 6 on valve B and the flow sensor B inlet. Take care not to overtighten these fingertight fittings.

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2. Rerun the system leak test.

• If the system leak test still fails, go to “To troubleshoot a leak in valve B.”

• If the system leak test passes, return the system to normal operation.

To troubleshoot a leak in valve B

1. Clean the rotor and stator in valve B as described in

“Cleaning the Rotor Seal and Stator” on page 74

.

2. If necessary, replace the rotor seal as described in

“Replacing the Rotor Seal” on page 77 .

Locating a Leak in the Solvent Path from Pump A to Flow Sensor A

The system leak test returned the following message (see Table 24 on page 221 ):

Check from pump A to flow sensor A for leaks. (Pump A flow loss >1000 nL/min).

The cause of this test result is a leak between flow sensor A and the inside of pump A. The system leak test script has detected that the piston movement is 1000 nL/min more than the flow measured at flow sensor A.

This procedure checks whether the leak is in the solvent path between the valve and the flow sensor. If the leak in not between the valve and the flow sensor, this procedure directs you to

the “Troubleshooting a Pump that Fails the Leaks Script” on page 216 where you determine if

the leak is coming from the solvent path between the pump and the valve or from the pump itself.

Figure 214

shows the solvent path from pump A to flow sensor A.

Figure 214. Solvent path from pump A to flow sensor A

Pump to pressure sensor Pressure sensor to valve A

Pump A

Valve to flow sensor

Valve A

2

3

1

6

4 5

Flow sensor A

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Troubleshooting the Results of the System Leak Test

To identify a leak in pump A or the solvent path between pump A and flow sensor A

1. Run the Leaks script for pump A only as follows: a. Set the parameters for the Leaks script as follows: i.

If you exited the Leaks script on the Maintenance > Scripts page, reopen it.

ii. Press the Parameters tab. iii. In the Value list, select A (pump A).

b. Press Start.

2. Depending on whether the leak test for pump A passes or fails, do the following:

• If the script passes, the leak is in the solvent path from valve A to flow sensor A.

Go to step 3

.

• If the script fails, the leak is in pump A. Go to “Troubleshooting a Pump that Fails the Leaks Script” on page 216 .

3. Check the connections to port 6 of valve A and the inlet to flow sensor A as follows:

• For the EASY-nLC II instrument, do the following:

– Hand tighten the PEEK fitting connected to the flow sensor inlet.

– Using a 1/4 in. open-ended wrench, tighten the stainless steel fitting connected to port 6 of valve A.

• For the EASY-nLC 1000 instrument, return the flow path upstream of valve S to atmospheric pressure as described in the

“Using nanoViper Fittings Quick Reference

Guide” on page 323 . Then individually disconnect and reconnect the nanoViper

fittings connected to port 6 on valve A and the flow sensor A inlet. Take care not to overtighten these fingertight fittings.

4. Rerun the system leak test. Depending on whether the test fails or passes, do the following:

• If the test fails, follow the instructions in “Maintaining the Rotary Valves” on page 74

to fix a leaking valve, which in this case is valve A. These instructions guide you through the process of cleaning the valve’s rotor seal and stator, rerunning the system leak test, and then replacing the rotor seal if necessary.

• If the test passes, the system is leak tight. You have completed this troubleshooting procedure. Return the EASY-nLC instrument to normal operation.

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Locating a Leak In or Between Valve S and Valve W

If the system leak test returns the following message (see Table 24 on page 221 ), follow the

workflow in

Figure 215 and

Figure 216

or the procedure on pages 228

– 234

to locate and remedy the leak.

Check from valve S to valve W, including valve W, for leaks.

Figure 215. Workflow for locating leaks in the solvent path between valves S and W or in these valves (Part 1)

System leak test fails and returns the following message:

Check from valve S to valve W, including valve W, for leaks.

For the EASY-nLC 1000, redo the nanoViper fitting connections to the leak test union (

step 1a on page 228

).

For the EASY-nLC II, hand tighten the fittings connected to the leak test union.

Start the Analytical Column Equilibration script ( step 1b on page 228

).

Done

No

Readback for flow sensor A

> 100 nL/min

Yes

Using a 1/4 in. wrench, tighten the blind, stainless steel fitting in port 1 of valve W ( step 2a on page 230

).

Done

No

Readback for flow sensor A

> 100 nL/min

Yes

Set valve W to the Center position ( step 3a on page 238

).

Service valve W ( page 74 ).

Then, rerun the system leak test (

page 218 ).

No

Readback for flow sensor A

> 100 nL/min

Yes

Go to

Figure 216

on the next page.

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Figure 216. Workflow for locating leaks in the solvent path between valves S and W or in these valves (Part 2)

Continued from

Figure 215 .

For the EASY-nLC 1000, redo the nanoViper connection to port 2 of valve W (

step 4a on page 230 ).

For the EASY-nLC II, use a 1/4 in. open-ended wrench to tighten the stainless steel fitting in port 2 of valve W.

Done

No

Readback for flow sensor A

> 100 nL/min

Yes

Disconnect the Waste In line from the leak test union (

step 5a on page 231 ).

Then, block the free end of the union with a blind fitting.

Install a new Waste In line.

Then, rerun the system leak test ( page 218

).

No

Readback for flow sensor A

> 100 nL/min

Yes

For the EASY-nLC 1000, redo the nanoViper connection to port 3 of valve S (

step 6a on page 232 ).

For the EASY-nLC II, use a 1/4 in. open-ended wrench tighten the stainless steel fitting in port 3 of valve S.

Reconnect the solvent lines and rerun the system leak test to confirm that the system is leak tight (

page 218

).

No

Readback for flow sensor A

> 100 nL/min

Yes

Disconnect the Column Out line from port 3 of valve S ( step 7a on page 233 ).

Then, block port 3 of valve S with a blind fitting.

Install a new Column Out line.

Then, rerun the system leak test ( page 218

).

No

Readback for flow sensor A

> 100 nL/min

Yes

Service valve S (

page 74 ).

Then, rerun the system leak test.

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Figure 217

shows the path between valves S and W for the EASY-nLC 1000 instrument.

Figure 217. Solvent path between valve S and valve W

Valve S

Valve W

Column Out line

Leak test union

Waste In line

Blind fitting connected to port 1

To locate and remedy a leak in valve S or valve W or in the solvent path between these valves

1. Make sure that the connections to the leak test union are secure as follows: a. Depending on the instrument model, do the following:

• For the EASY-nLC II instrument, hand tighten the PEEK fittings.

• For the EASY-nLC 1000 instrument, do the following: i.

Disconnect the nanoViper fittings from the union.

ii. Reinsert the nanoViper fittings into the stainless steel union. Then, slowly tighten the fittings until you feel resistance. iii. Tighten both fittings by an additional 45 degrees (1/8-turn). Do not tighten the fittings by more than 90 degrees (1/4-turn).

For more information, see “Using nanoViper Fittings” on page 83 .

.

Note The connections to the leak test union are one of the most common sources of leaks for the system leak test.

b. For leak testing, set up and start the Analytical Column Equilibration script as follows: i.

On the Maintenance > Scripts page, select Prepare in the Category list, and then select Analytical Col Equilibration in the Name list.

ii. Press the Parameters tab.

iii. In the Volume [μL] box, enter 140. This volume should be sufficient to keep the solvent flow on during this troubleshooting procedure. iv. Leave the Flow box [μL/min] box empty so that pump A operates at the set pressure.

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Troubleshooting the Results of the System Leak Test v.

In the Max Pressure [bar] box, enter 280 bar for the EASY-nLC II instrument or 980 bar for the EASY-nLC 1000 instrument.

vi. Press Start.

The instrument places the valves in the following positions, and pump A begins pumping solvent through flow sensor A.

• Valve A: 1–6

• Valve B: Center

• Valve S: 1–2

• Valve W: 1–2

Figure 218

shows the solvent system schematic on the Home > Overview page. Use this page to monitor the readback for flow sensor A and to access the valve controls.

For information about changing the valve positions, see

“Using the Valve Controls” on page 204

.

Figure 218. Home > Overview page showing the readback for flow sensor A

Readback for flow sensor A with a flow < 100 nL/min

Press this icon to open the controls for valve W.

Thermo Scientific c.

On the Home > Overview page, check the readback for flow sensor A. Then, depending on the flow rate, do one of the following:

• If the flow rate is <100 nL/min, you have remedied the leak and completed this troubleshooting procedure. Return the instrument to normal operation.

• If the flow rate is >100 nL/min, go to the next step.

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2. Make sure that the connection between the blind fitting and port 1 of valve W is leak tight as follows: a. Using a 1/4 in. open-ended wrench, tighten the stainless steel blind fitting connected to port 1 of valve W.

b. With the Analytical Column Equilibration script running, monitor flow sensor A.

Then, depending on the flow rate, do one of the following:

• If the flow is <100 nL/min, you have remedied the leak and completed this troubleshooting procedure. Return the instrument to normal operation.

• If the flow is >100 nL/min, go to the next step.

3. Check if valve W is leaking as follows: a. On the Home > Overview page, press the valve W icon and set the valve to the

Center position.

Placing valve W in the Center position blocks the solvent flow into valve W and eliminates valve W as the possible source of the leak. b. With the Analytical Column Equilibration script running, monitor flow sensor A.

Depending on the flow rate, do one of the following:

• If the flow is <100 nL/min with valve W removed from the solvent path, valve W is the source of the system leak. To fix the leaking valve, follow the instructions in

“Maintaining the Rotary Valves” on page 74 to clean the valve, replace the rotor,

and if necessary, replace the stator. Then, rerun the system leak test.

• If the flow is >100 nL/min, go to the next step.

4. Check for a leak-tight connection to port 2 of valve W as follows: a. Depending on the instrument model, do the following:

• For the EASY-nLC II instrument, use a 1/4 in. open-ended wrench to tighten the fitting to port 2 of valve W.

• For the EASY-nLC 1000 instrument, do the following: i.

On the Home > Overview page, change the position of the following valves to return the system to atmospheric pressure:

– Valve S: Center

– Valve W: 1–6 ii. Disconnect and then reconnect the nanoViper fitting from port 2 of valve W. Take care to avoid overtightening the fingertight fitting.

iii. On the Home > Overview page, reset valves A and S as follows:

– Valve S: 1–2

– Valve W: 1–2

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Troubleshooting the Results of the System Leak Test b. With the Analytical Column Equilibration script running, monitor flow sensor A.

Then, depending on the flow rate, do one of the following:

• If the flow is <100 nL/min, you have remedied the leak and completed this troubleshooting procedure. Return the instrument to normal operation.

• If the flow is >100 nL/min, go to the next step.

5. Check the Waste In line as follows: a. Depending on the instrument model, do the following:

• For the EASY-nLC II instrument, disconnect the Waste In line from the leak test union. Then, block the free end of the union with a blind fitting.

• For the EASY-nLC 1000 instrument, do the following: i.

On the Home > Overview page, change the position of the following valves to return the system to atmospheric pressure:

– Valve S: Center

– Valve W: 1–6 ii. Using the black knurled nut to loosen the nanoViper fitting, disconnect the Waste In line from the leak test union. Then, block the free end of the union with a blind fitting.

iii. On the Home > Overview page, reset valves S and W as follows:

– Valve S: 1–2

– Valve W: 1–2

Figure 219

shows the Waste In line disconnected from the leak test union and the free end of the union plugged with a blind fitting.

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Troubleshooting the Results of the System Leak Test

Figure 219. Setup for eliminating the Waste In line as the source of the system leak

Blind fitting

Valve S Valve W

From the mixing Tee b. With the Analytical Column Equilibration script running, monitor flow sensor A.

Then, depending on the flow rate, do one of the following:

• If the flow is <100 nL/min, install a new Waste In line. Then, reconnect the

solvent lines and rerun the system leak test as described in “Running a System

Leak Test” on page 218

.

For ordering information, see Appendix C, “Consumables and Replacement

Parts.”

• If the flow is >100 nL/min, go to the next step.

6. Check for a leak-tight connection to port 3 of valve S as follows: a. Depending on the instrument model, do the following:

• For the EASY-nLC II instrument, use a 1/4 in. open-ended wrench to tighten the fitting to port 3 of valve S.

• For the EASY-nLC 1000 instrument, do the following: i.

On the Home > Overview page, set valve S to the Center position to return the system to atmospheric pressure: ii. Disconnect and then reconnect the Column Out line to port 3 of valve S.

Take care to avoid overtightening the fingertight fitting.

iii. On the Home > Overview page, reset valve S to position 1–2.

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Troubleshooting

Troubleshooting the Results of the System Leak Test b. With the Analytical Column Equilibration script running, monitor flow sensor A.

Then, depending on the flow rate, do one of the following:

• If the flow is <100 nL/min, reconnect the solvent lines and rerun the system leak

test as described in “Running a System Leak Test” on page 218

to confirm that the system is leak tight.

• If the flow is >100 nL/min, go to the next step.

7. Determine whether the leak is in the Column Out line or in valve S as follows: a. Depending on the instrument model, do the following:

• For the EASY-nLC II instrument, use a 1/4 in. open-ended wrench to disconnect the Column Out line from port 3 of valve S. Plug the port with a blind fitting.

Then, use a 1/4 in. open-ended wrench to tighten the fitting.

• For the EASY-nLC 1000 instrument, do the following: i.

On the Home > Overview page, set valve S to the Center position to return the system to atmospheric pressure: ii. Using the black knurled nut, disconnect the Column Out line from port 3 of valve S. Then, block port 3 of valve S with a blind fitting.

iii. On the Home > Overview page, reset valve S to position 1–2.

Figure 220

shows port 3 of valve S plugged with a blind fitting.

Figure 220. Port 3 of valve S plugged with a blind fitting

Valve S Valve W

Blind fitting in port 3 of valve S

From the mixing Tee

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Troubleshooting

Troubleshooting the Results of the System Leak Test b. With the Analytical Column Equilibration script running, monitor flow sensor A.

Then, depending on the flow rate, do one of the following:

• If the flow is <100 nL/min, install a new Column Out line. Then, reconnect the

solvent lines and rerun the system leak test as described in “Running a System

Leak Test” on page 218

.

For ordering information, see Appendix C, “Consumables and Replacement

Parts.”

• If the flow is >100 nL/min, remedy the leak in valve S as described in

“Maintaining the Rotary Valves” on page 74

. Then, rerun the system leak test as described in

“Running a System Leak Test” on page 218

.

Locating a Leak Between the Flow Sensors and Valve S

When one or more of the connections are leaking in the solvent path between the flow sensor

outlets and the mixing Tee inlets, the system leak test returns this message (see Table 24

on

page 221 :

Check solvent lines between flow sensor A and valve S, including the mixing Tee and flow sensor B, for leaks.

As the flow sensors and mixing Tee differ for the EASY-nLC 1000 and EASY-nLC II instruments, follow the topic for your instrument to locate and remedy the leak:

“EASY-nLC 1000: Locating a Leak Between the Flow Sensors and Valve S,”

on pages

235

– 245

“EASY-nLC II: Locating a Leak Between the Flow Sensors and Valve S” on page 246

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Troubleshooting the Results of the System Leak Test

EASY-nLC 1000: Locating a Leak Between the Flow Sensors and Valve S

To determine whether the leak is coming from the connection to port 4 of valve S, the connection to the mixing Tee outlet, or the connections between the flow sensor

outlets and the mixing Tee inlets, follow the workflow in Figure 221

or the procedure on

pages 236

– 239

.

Figure 221. EASY-nLC 1000 workflow for locating leaks between the flow sensors and valve S

System leak test fails and returns the following message:

Check solvent lines between flow sensor A and valve S, including the mixing Tee and flow sensor B, for leaks.

Redo the nanoViper fitting connection to port 4 of valve S (

step 1a on page 236 ).

Start the Analytical Column Equilibration script (

step 1b on page 236

).

Done

No

Readback for flow sensor A

> 100 nL/min

Yes

Disconnect the nanoViper fitting from the mixing Tee outlet.

Then, block the mixing Tee outlet with a blind fitting ( step 2 on page 236

).

Install a new solvent line between port 4 of valve S and the mixing Tee outlet

(

step 2d on page 238 ).

Then, reconnect the plumbing and rerun the system leak test (

page 218 ).

No

Readback for flow sensor A

> 100 nL/min

Yes

Disconnect the blind fitting from the mixing Tee outlet, and then reconnect the solvent line to the mixing Tee outlet. Disconnect the solvent line from the mixing

Tee inlet for the solvent B channel, and then block the inlet with a blind fitting

( step 3 on page 238

).

No

Go to “EASY-nLC 1000: Locating a Leak Between

Flow Sensor A and the Mixing Tee” on page 240 .

Readback for flow sensor A

> 100 nL/min

Yes

Go to “EASY-nLC 1000: Locating a Leak Between

Flow Sensor B and the Mixing Tee” on page 243 .

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Troubleshooting

Troubleshooting the Results of the System Leak Test

To locate leaks between the flow sensors and valve S in the EASY-nLC 1000 instrument

1. To ensure a leak-tight connection to port 4 of valve S, do the following: a. Disconnect and then reconnect the nanoViper fitting to port 4 of valve S.

b. From the Maintenance > Scripts page, start the Analytical Column Equilibration script with the maximum pressure set to 980 bar and the volume set to 140 μL.

For information about setting up the Analytical Column Equilibration script, see

step 1b on page 228 .

When you start the Analytical Column Equilibration script, the instrument sets the rotary valves to these positions and begins pumping solvent through flow sensor A:

• Valve A: 1–6

• Valve B: Center

• Valve S: 1–2

• Valve W: 1–2 c.

On the Home > Overview page, check the readback for flow sensor A. Then, depending on the flow rate, do one of the following:

• If the flow rate is <100 nL/min, you have remedied the leak and completed this troubleshooting procedure. Return the instrument to normal operation.

• If the flow rate is >100 nL/min, go to the next step.

2. To check the solvent line that connects the mixing Tee outlet to port 4 of valve S, do the following: a. On the Home > Overview page, set valves A and W to the following positions to return the system to atmospheric pressure:

• Valve A: Center

• Valve W: 1–6

For information about changing the valve positions, see

“Using the Valve Controls” on page 204

.

b. Disconnect the nanoViper fitting from the mixing Tee outlet.Then, block the mixing

Tee outlet with a blind fitting (see

Figure 222 ).

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Figure 222. Mixing Tee outlet plugged with a blind fitting

Waste In line

Leak test union

Column Out line

Solvent line that connects the mixing Tee outlet to port 4 of valve S

Inline filter connected to the outlet of flow sensor A

Blind fitting connected to the mixing Tee outlet

Inline filter connected to the outlet of flow sensor B c.

On the Home > Overview page, reset these valve positions:

• Valve A: 1–6

• Valve W: 1–2

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Troubleshooting

Troubleshooting the Results of the System Leak Test d. On the Home > Overview page, check the readback for flow sensor A. Then, depending on the flow rate, do one of the following:

• If the flow rate is <100 nL/min, do the following: i.

Stop the Analytical Column Equilibration script and return the system to atmospheric pressure.

ii. Disconnect the damaged solvent line from port 4 of valve S.

iii. Install a new solvent line to connect the mixing Tee outlet to port 4 of valve S.

For ordering information, see Table 33 on page 316

.

iv. Rerun the system leak test (see

“Running a System Leak Test” on page 218 ).

• If the flow rate is >100 nL/min, go to the next step.

3. To eliminate the solvent B channel as the leak source, do the following: a. Set valve A to the Center position to return the system to atmospheric pressure while maintaining pressure on the front end of the system. b. Disconnect the blind fitting from the mixing Tee outlet, and then reconnect the solvent line to the mixing Tee outlet.

c.

Disconnect the solvent line from the mixing Tee inlet for the solvent B channel, and then block the inlet with a blind fitting.

Figure 223

shows the solvent line disconnected from the mixing Tee and a blind fitting in the inlet port of the mixing Tee for the solvent B channel.

d. Reset valve A to position 1–6.

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Figure 223. Mixing Tee inlet for the solvent B channel plugged with a blind fitting

Leak test union

6

Troubleshooting

Troubleshooting the Results of the System Leak Test

Thermo Scientific

Solvent line that connects the mixing Tee outlet to port 4 of valve S

Mixing Tee inlet for the solvent B channel plugged with a blind fitting e.

On the Home > Overview page, check the readback for flow sensor A. Then, depending on the flow rate, do one of the following:

• If the flow rate is <100 nL/min, go to

“EASY-nLC 1000: Locating a Leak

Between Flow Sensor B and the Mixing Tee” on page 243 , as removing the

solvent B channel from the flow path remedied the leak.

• If the flow rate is >100 nL/min, the leak is in the solvent A channel, as removing the solvent B channel from the solvent path did not remedy the leak. Go to the

next procedure, “EASY-nLC 1000: Locating a Leak Between Flow Sensor A and the Mixing Tee.”

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EASY-nLC 1000: Locating a Leak Between Flow Sensor A and the Mixing Tee

By following the workflow in

Figure 221 on page 235

or the procedure on pages 236

– 239

, you determined that the leaking connection is between flow sensor A and the mixing Tee. The

system plumbing is set up as shown in Figure 223 on page 239 , with the solvent channel B

disconnected from the mixing Tee.

Follow the workflow in

Figure 224

or the procedure on pages 241 –

242 to locate and remedy

the leak.

Figure 224. EASY-nLC 1000 workflow for locating a leak between flow sensor A and the mixing Tee (Part 1)

You determined that the leak is in the solvent path between the outlet of flow sensor A and the mixing Tee inlet.

Disconnect the solvent line from the outlet of flow sensor A.

Then, block the outlet with a blind fitting ( step 1b on page 241

).

Install a new solvent line between the flow sensor A outlet and the mixing Tee inlet (

step 1d on page 242 ).

Then, reconnect all of the solvent lines and rerun the system leak test (

page 218

).

No

Readback for flow sensor A

> 100 nL/min

Yes

Using a 5/16 in. open-ended wrench, tighten the inline filter to the outlet of flow sensor A (

step 2a on page 242

).

Reconnect all of the solvent lines, and then rerun the system leak test (

page 218 ).

No

Readback for flow sensor A

> 100 nL/min

Yes

Replace the inline filter (

page 81

). Then, reconnect all of the solvent lines and

rerun the system leak test ( page 218

).

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To locate and remedy a leak between flow sensor A and the mixing Tee in the

EASY-nLC 1000

1. To check if you must replace the solvent line that connects the outlet of flow sensor A to the mixing Tee, do the following: a. Set valve A to the Center position.

b. Disconnect the solvent line from the inline filter that connects to the outlet of flow sensor A. Then, block the inline filter with a blind fitting.

Figure 225

shows the solvent path blocked at the outlet of flow sensor A.

c.

Reset valve A to position 1–6.

Figure 225. Solvent A channel blocked at the outlet of flow sensor A

If the flow is <100 nL/min with this line removed from the solvent path, install a new solvent line.

Outlet of flow sensor A plugged with a blind fitting

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Troubleshooting

Troubleshooting the Results of the System Leak Test d. Check the readback for flow sensor A. Then, depending on the flow rate, do one of the following:

• If the flow rate is <100 nL/min, do the following: i.

Stop the Analytical Column Equilibration script.

ii. Disconnect the damaged solvent line from the mixing Tee’s inlet for the solvent A channel.

iii. Install a new solvent line between the mixing Tee’s inlet for the solvent A channel and the flow sensor’s outlet.

For ordering information, see Table 33 on page 316

.

iv. Reconnect the solvent lines and rerun the system leak test (see “Running a

System Leak Test” on page 218

).

• If the flow rate is >100 nL/min, go to the next step.

2. To check the connection between the inline filter and the flow sensor A outlet, do the following: a. Using a 5/16 in. open-ended wrench, tighten the inline filter to the outlet of flow sensor A. b. Check the readback for flow sensor A. Then, depending on the flow rate, do one of the following:

• If the flow rate is <100 nL/min, do the following: i.

Stop the Analytical Column Equilibration script.

ii. Reconnect all of the solvent lines.

iii. Rerun the system leak test (see

“Running a System Leak Test” on page 218 ).

• If the flow rate is >100 nL/min, do the following: i.

Stop the Analytical Column Equilibration script.

ii. Remove the damaged inline filter and install a new inline filter.

iii. Rerun the system leak test (see

“Running a System Leak Test” on page 218 ).

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Troubleshooting the Results of the System Leak Test

EASY-nLC 1000: Locating a Leak Between Flow Sensor B and the Mixing Tee

By following the workflow in

Figure 221 on page 235

or the procedure on pages 236

– 239

, you determined that the leaking connection is between the flow sensor B outlet and the mixing Tee inlet. The system plumbing is set up as shown in

Figure 223 on page 239

, with the solvent channel B disconnected from the mixing Tee inlet.

Follow the workflow in Figure 226

or the procedure on pages

244 –

245 to locate and remedy

the leak.

Figure 226. EASY-nLC 1000 workflow for locating a leak between flow sensor B and the mixing Tee

You determined that the leak is in the solvent path between the outlet of flow sensor B and the inlet of the mixing Tee.

Remove the blind fitting from the solvent B channel inlet of the mixing Tee. Reconnect the solvent line to the solvent B channel inlet of the mixing Tee. Redo the nanoViper connection to the outlet of flow sensor B (

step 1 on page 244 ).

The system plumbing now matches that for a system leak test (see Figure 35 on page 45

).

Done

No

Readback for flow sensor A

> 100 nL/min

Yes

Using a 5/16 in. open-ended wrench, tighten the inline filter to the outlet of

flow sensor B ( step 2 on page 244 ).

Done

No

Readback for flow sensor A

> 100 nL/min

Yes

Disconnect the solvent line from the outlet of flow sensor B. Then, plug the free end of

the solvent line with a leak test union and a blind fitting ( step 3 on page 244

).

Install a new inline filter (

page 81

Reconnect all of the solvent lines. Then,

rerun the system leak test (

).

page 218

).

No

Readback for flow sensor A

> 100 nL/min

Yes

Install a new solvent line between the flow sensor B outlet and the mixing Tee inlet

( step 3d on page 245

). Rerun the system leak test (

page 218 ).

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To locate a leak between flow sensor B and the mixing Tee in the EASY-nLC 1000

1. To ensure leak-tight connections for the solvent line that connects the outlet of flow sensor B to the mixing Tee inlet, do the following: a. Set valve A to the Center position.

b. Set up the system plumbing as follows: i.

Remove the blind fitting from the mixing Tee’s inlet for the solvent B channel. ii. Reconnect the solvent line to the solvent B channel inlet of the mixing Tee.

iii. Redo the nanoViper connection to the mixing Tee’s inlet for the solvent B channel.

The system plumbing now matches that for a system leak test.

c.

Reset valve A to position 1–6.

d. Check the readback for flow sensor A. Then, depending on the flow rate, do one of the following:

• If the flow rate is <100 nL/min, you have completed this troubleshooting procedure. Return the instrument to normal operation.

• If the flow rate is >100 nL/min, go to the next step.

2. Using a 5/16 in. open-ended wrench, tighten the inline filter to the outlet of flow sensor B.

3. To determine whether the solvent line or the inline filter is the leak source, do the following: a. Set valve A to the Center position.

b. Disconnect the solvent line from the outlet of flow sensor B. Then, plug the free end

of the solvent line with a leak test union and a blind fitting (see Figure 227

).

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Figure 227. Solvent line disconnected from the outlet of flow sensor B

Inlets of the EASY-nLC 1000 flow sensors

Thermo Scientific

Inline filter connected to the outlet of flow sensor B

Solvent line disconnected from the outlet of flow sensor B and plugged with a union and a blind fitting c.

Reset valve A to position 1–6.

d. Check the readback for flow sensor A. Then, depending on the flow rate, do one of the following:

• If the flow rate is <100 nL/min, do the following: i.

Stop the Analytical Column Equilibration script.

ii. Remove the damaged inline filter from the outlet of flow sensor B and install a new inline filter.

For ordering information, see Table 35 on page 319

.

iii. Reconnect the system plumbing and rerun the system leak test (see

“Running a System Leak Test” on page 218

).

• If the flow rate is >100 nL/min, do the following: i.

Stop the Analytical Column Equilibration script.

ii. Install a new solvent line between the flow sensor B outlet and the mixing

Tee inlet.

For ordering information, see Table 33 on page 316

.

iii. Rerun the system leak test (see

“Running a System Leak Test” on page 218 ).

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Troubleshooting the Results of the System Leak Test

EASY-nLC II: Locating a Leak Between the Flow Sensors and Valve S

Figure 228 on page 247 shows the solvent lines that connect the flow sensor outlets to the

mixing Tee and the solvent line that connects the mixing Tee outlet to port 4 of valve S.

To systematically locate and remedy a leak in an EASY-nLC II instrument from the flow sensor outlets to port 4 of valve S

1. To ensure a leak-tight connection to port 4 of valve S, do the following: a. Tighten the fitting in port 4 of valve S.

b. From the Maintenance > Scripts page, start the Analytical Column Equilibration script with the maximum pressure set to 280 bar and the volume set to 140 μL.

For information about setting up the Analytical Column Equilibration script, see

step 1b on page 228 .

c.

On the Home > Overview page, check the readback for flow sensor A. Then, depending on the flow rate, do one of the following:

• If the flow rate is <100 nL/min, you have remedied the leak and completed this troubleshooting procedure. Return the instrument to normal operation.

• If the flow rate is >100 nL/min, go to the next step.

2. Hand tighten the following PEEK fittings:

• Fittings connected to the mixing Tee

• Fittings connected to the flow sensor outlets

3. With the Analytical Column Equilibration script running, monitor flow sensor A.

Depending on the flow rate, do one of the following:

• If the flow is <100 nL/min, tightening the solvent line connections remedied the leak, and you have completed this troubleshooting procedure. Return the instrument to normal operation.

• If the flow is >100 nL/min, the leak is coming from one of the solvent lines. Go to the next step.

Note For information about ordering solvent lines for the EASY-nLC II instrument,

see Table 32 on page 314

.

4. Replace the solvent line that connects the mixing Tee outlet and port 4 of valve S. Restart the Analytical Column Equilibration script and monitor flow sensor A.

• If the flow is <100 nL/min, you have completed this troubleshooting procedure.

• If the flow is >100 nL/min, go to the next step.

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5. Replace the solvent line that connects the outlet of flow sensor A to the mixing Tee inlet.

Restart the Analytical Column Equilibration script and monitor flow sensor A.

• If the flow is <100 nL/min, you have completed this troubleshooting procedure.

• If the flow is >100 nL/min, go to the next step.

6. Replace the solvent line that connects the outlet of flow sensor B to the mixing Tee inlet.

Restart the Analytical Column Equilibration script and monitor flow sensor A.

• If the flow is <100 nL/min, you have completed this troubleshooting procedure.

• If the flow is >100 nL/min, contact your local Thermo Fisher Scientific field service engineer.

Figure 228. Solvent lines that connect the flow sensor outlets to the mixing Tee and the solvent line that connects the mixing

Tee outlet to port 4 of valve S in the EASY-nLC II instrument

Solvent line that connects the outlet of flow sensor A to the mixing Tee

Mixing Tee with fingertight connections

Column Out and

Waste In lines connected to the leak test union

Valve W

3

2

4

1

6

5

Solvent line that connects the outlet of flow sensor B to the mixing Tee

Valve S

3

2

4

1

6

5

Solvent line that connects the mixing Tee outlet to port 4 of valve S

Flow sensor outlets with fingertight connections

+

Solvent lines that connect valves A and B to the flow sensor inlets

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Identifying a Leaking Check Valve

Identifying a Leaking Check Valve

The EASY-nLC instrument has four check valves. Each solvent inlet line connects to a check valve assembly with a solvent-side check valve and a waste-side check valve.

CAUTION Wear powder-free gloves and safety glasses when handling parts of the LC system that come into contact with solvents.

To determine if the waste-side check valve is leaking

1. Run the Purge Solvent script as follows: a. Press Maintenance > Scripts. b. In the Category list, select Prepare. c.

In the Name list, select Purge Solvent. d. Press the Parameters tab.

e.

Select the check box for the appropriate pump.

f.

Press Start.

2. As the pump is filling, observe the waste line.

If you observe solvent moving toward the check valve assembly, the waste-side check valve is leaking.

3. If the waste-side check valve is leaking, replace it and repeat the Purge Solvent script.

To determine if the solvent-side check valve is leaking

1. Start the Purge Solvent script.

2. As the pump is emptying, observe the solvent line.

If you observe solvent moving toward the solvent bottle from the check valve assembly, the solvent-side check valve is leaking.

3. If the solvent-side check valve is leaking, replace it and repeat the Purge Solvent script.

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Troubleshooting a System Blockage

Troubleshooting a System Blockage

Use this troubleshooting procedure when the back pressure for a chromatographic method rises above the expected level or when the flow decreases below the set rate for the column equilibration or sample loading steps.

In most cases, an increase in back pressure is caused by a blocked column or a blocked

Column Out line. The Column Out line has an ID of 30 μm for the EASY-nLC II instrument and 20 μm for the EASY-nLC 1000 instrument and is the smallest ID line that the sample passes through, from sample pickup to the columns.

Troubleshooting a system blockage requires these tools and materials.

Tools

1/4 in. open-ended wrench to loosen the stainless steel fittings

Parts and materials

• Powder-free gloves

• EASY-nLC II instrument: HPLC union, P/N SC600

• EASY-nLC 1000 instrument: HPLC union, P/N SC900

CAUTION Wear powder-free gloves and safety glasses when handling parts of the LC system that come into contact with solvents.

When running the Back Pressure script, the system also checks for an unusually low back pressure. If the back pressure is unusually low, the system probably has a leak.

To troubleshoot a system blockage, follow these procedures:

1.

“Running the Back Pressure Test for the A and B Solvent Paths” on page 250

2. Depending on the test results, replace the blocked components in the column assembly or follow one or both of these procedures:

“Troubleshooting a System Blockage when Test Solvent A Fails” on page 251

“Troubleshooting a System Blockage when Test Solvent B Fails” on page 257

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Troubleshooting

Troubleshooting a System Blockage

Running the Back Pressure Test for the A and B Solvent Paths

The Back Pressure script determines instrument back pressure for solvent lines A and B. The script runs at a preset flow and measures the back pressure on the system.

IMPORTANT Before you perform the Back Pressure script, ensure that the solvent A bottle contains water and the solvent B bottle contains acetonitrile. This test is not valid for other solvents.

To run the back pressure script for both the A and B solvent paths

1. Remove the columns and connect the Column Out line to the Waste In line using the appropriate HPLC union.

Instrument

EASY-nLC II

EASY-nLC 1000

Union

SC600

SC900

2. Run the Back Pressure scripts for both Solvent A and Solvent B as follows: a. Press Maintenance > Scripts.

b. In the Category list, select Test.

c.

In the Name list, select Back Pressure.

d. Press the Parameters tab.

e.

Select the Test Solvent A and Test Solvent B check boxes. f.

Press Start.

3. Depending on the test results, do one of the following:

• If the Test Solvent A test fails and the following message appears, go to

“Troubleshooting a System Blockage when Test Solvent A Fails” on page 251 .

– EASY-nLC II instrument—Pressure is xxx bar (exceeds 100 bar limit), test failed!

– EASY-nLC 1000 instrument—Pressure is xxx bar (exceeds 175 bar limit), test failed!

• If the Test Solvent B test fails and the following message appears, go to

“Troubleshooting a System Blockage when Test Solvent B Fails” on page 257 .

Pressure is xxx bar (exceeds 100 bar limit), test failed!

• If both Test Solvent A and Test Solvent B tests pass and the following message appears, inspect and if necessary replace the columns, the packed emitter, or both.

Pressure is xxx bar, test passed.

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Troubleshooting a System Blockage

Troubleshooting a System Blockage when Test Solvent A Fails

Follow this procedure when the Back Pressure test for Test Solvent A fails (see

“Running the

Back Pressure Test for the A and B Solvent Paths” on page 250

).

To verify the system blockage

1. Run the Precolumn Equilibration script and check the back pressure as follows: a. On the Maintenance > Scripts page, select Prepare in the Category list, and then select Precolumn Equil in the Name list.

b. Press the Parameters tab.

c.

In the Volume [μL] box, enter 140 μL.

d. In the Flow [μL/min] box, enter 2 μL/min.

e.

Leave the Max Pressure [bar] box blank.

f.

Press Start.

g. Monitor the pump A pressure trace.

2. Depending on the back pressure, do the following:

• If the back pressure is below the pressure threshold, the system blockage has been removed. To confirm that the back pressure is below the pressure threshold, rerun the

Back Pressure script with the Test Solvent A check box selected.

• If the back pressure is above the pressure threshold, keep the Precolumn Equil script running and go to

“To locate the source of the blockage when the Test Solvent A test fails” on page 253 .

Instrument

EASY-nLC II

EASY-nLC 1000

Pressure threshold

100 bar

175 bar

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Troubleshooting a System Blockage

Figure 229

shows the areas that the following procedure systematically checks for blockage.

Figure 229. Solvent path from pressure sensor A to the Waste In line

2.

Waste In line

2

3

1

6

4 5

3.

Column Out line

4.

Valve S

2

3

1

6

4 5

5.

Valve S (port 4) to mixing Tee

6.

Mixing Tee

8.

Inline filter

(EASY-nLC 1000 instrument)

10.

Inline filter

(EASY-nLC 1000 instrument)

7.

Mixing Tee to flow sensor A outlet

9.

Flow sensor A

12.

Valve A

11.

Valve A (port 6) to flow sensor inlet

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1.

Valve W

Note.

Disconnect the Column Out and the

Waste In lines from the union after testing valve W for blockage.

2

3

1

6

4 5

13. Valve A (port 1) to pressure sensor

Thermo Scientific

6

Troubleshooting

Troubleshooting a System Blockage

To locate the source of the blockage when the Test Solvent A test fails

1. To determine if valve W is blocked, do the following: a. Make sure that the Precolumn Equilibration script is set up and running as described in

step 1 on page 251

and that the Column Out and Waste In lines are connected with a union.

b. Disconnect the Waste In line from port 2 of valve W (see Figure 229 on page 252 ).

c.

Depending on the back pressure, do one of the following:

• If the pressure is below the threshold, valve W is blocked. Go to “Maintaining the Rotary Valves” on page 74

.

• If the pressure is above the threshold, go to

step 2

.

2. To determine if the Waste In line is blocked, do the following: a. Disconnect the Column Out line and the Waste In line from the union.

Thermo Scientific

Column Out line Union Waste In line b. Depending on the back pressure, do one of the following:

• If the pressure is below the threshold, the Waste In line is blocked. Discontinue this troubleshooting procedure and replace the Waste In line with a new line.

Instrument

EASY-nLC II

EASY-nLC 1000

Pressure threshold

100 bar

125 bar

• If the pressure is above the threshold, go to

step 3

.

3. To determine if the Column Out line is blocked, do the following: a. Disconnect the Column Out line from port 3 of valve S. b. Depending on the back pressure, do one of the following:

• If pressure is below the threshold, the Column Out line is blocked. Discontinue this troubleshooting procedure and replace the Column Out line with a new line.

Instrument

EASY-nLC II

EASY-nLC 1000

Pressure threshold

100 bar

90 bar

• If the pressure is above the threshold, reconnect the Column Out line, and then go to

step 4

.

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Troubleshooting a System Blockage

4. To determine if valve S is partially or totally blocked, do the following: a. Disconnect the mixing Tee line from port 4 of valve S. b. Depending on the back pressure, do one of the following:

• If the pressure is below the threshold, valve S is partially or totally blocked.

Discontinue this troubleshooting procedure and go to

“Maintaining the Rotary

Valves” on page 74 .

Instrument

EASY-nLC II

EASY-nLC 1000

Pressure threshold

100 bar

90 bar

• If the pressure is above the threshold, reconnect the mixing Tee line to port 4 of

valve S, and then go to step 5 .

5. To determine if the line that connects valve S to the mixing Tee is blocked, do the following: a. Disconnect the line from the mixing Tee.

b. Depending on the back pressure, do one of the following:

• If the pressure is below the threshold, the line that connects the mixing Tee to valve S is blocked. Replace it.

Instrument

EASY-nLC II

EASY-nLC 1000

Pressure threshold

100 bar

65 bar

• If the pressure is above the threshold, reconnect the line to the mixing Tee, and

then go to step 6

.

6. To determine if the mixing Tee is blocked, do the following: a. Disconnect the line that exits flow sensor A from the mixing Tee.

b. Depending on the back pressure, do one of the following:

• If the pressure is below the threshold, the mixing Tee is blocked. Clean the mixing Tee, or if necessary replace it.

Instrument

EASY-nLC II

EASY-nLC 1000

Pressure threshold

100 bar

65 bar

• If the pressure is above the threshold, reconnect the line to the mixing Tee, and

then go to step 7

.

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Troubleshooting

Troubleshooting a System Blockage

7. To determine if the line that connects the mixing Tee to flow sensor A is blocked, do the following: a. Disconnect the line from the outlet of flow sensor A.

b. Depending on the back pressure, do one of the following:

• If the pressure is <20 bar, the line that connects the mixing Tee to flow sensor A is blocked. Replace it with a new solvent line.

• If the pressure is >20 bar, reconnect the line to flow sensor A, and then go to

step 8 for an EASY-nLC 1000 instrument or step 9

for an EASY-nLC II instrument.

8. To determine if the inline filter connected to the flow sensor outlet of the

EASY-nLC 1000 system is blocked, do the following: a. Remove the inline filter from the flow sensor outlet.

b. Depending on the back pressure, do one of the following:

• If the pressure is <20 bar, the inline filter is blocked. Replace it with a new filter.

• If the pressure is >20 bar, reconnect the inline filter to flow sensor A, and then go

to step 9 .

9. To determine if flow sensor A is blocked, do the following: a. Disconnect the solvent line from the flow sensor A inlet.

b. Depending on the back pressure for flow sensor A, do one of the following:

• If the pressure is <20 bar, flow sensor A is blocked. Replace it with a new flow

sensor as described in “Replacing a Flow Sensor” on page 92 .

• If the pressure is >20 bar, reconnect the line to flow sensor A, and then go to

step 10 for an EASY-nLC 1000 system or

step 11

for an EASY-nLC II system.

10. To determine if the inline filter connected to the flow sensor inlet of the EASY-nLC 1000 system is blocked, do the following: a. Remove the inline filter from the flow sensor inlet.

b. Depending on the back pressure, do one of the following:

• If the pressure is <20 bar, the inline filter is blocked. Replace it with a new inline filter.

• If the pressure is >20 bar, reconnect the inline filter to flow sensor A, and then go

to step 11

.

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Troubleshooting a System Blockage

11. To determine if the line that connects valve A to flow sensor A is blocked, do the following: a. Disconnect the solvent line from port 6 of valve A.

b. Depending on the back pressure, do one of the following:

• If the pressure is <20 bar, the line from valve A to flow sensor A is blocked.

Replace it with a new solvent line.

• If the pressure is >20 bar, reconnect the line to valve A, and then go to step 12

.

12. To determine if valve A is blocked, do the following: a. Disconnect the solvent line from port 1 of valve A.

b. Depending on the back pressure, do one of the following:

• If the pressure is <20 bar, valve A is blocked. Inspect the valve and clean the stator. If the blockage persists, replace the rotor seal.

• If the pressure is >20 bar, reconnect the line to valve A, and then go to step 13

.

13. To determine if the line from pressure sensor A to valve A is blocked or if pressure sensor A is blocked, do the following: a. Disconnect the solvent line from the outlet of pressure sensor A.

b. Depending on the back pressure, do one of the following:

• If the pressure is <20 bar, the line that connects pressure sensor A to valve A is blocked. Replace it with a new solvent line.

• If the pressure is >20 bar, pressure sensor A is blocked. Replace it with a new pressure sensor as described in

“Replacing a Pressure Sensor for the PLU Pump” on page 90

.

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Troubleshooting

Troubleshooting a System Blockage

Troubleshooting a System Blockage when Test Solvent B Fails

Follow this procedure when the Back Pressure test for Test Solvent B fails (see “To run the back pressure script for both the A and B solvent paths” on page 250

).

Figure 230

shows the areas that this procedure systematically checks for blockage.

Figure 230. Solvent system from pressure sensor B to the mixing Tee

1.

Mixing Tee

2.

Flow sensor outlet to the mixing Tee

3.

Inline filter for the

EASY-nLC 1000 instrument

4.

Flow sensor

5.

Inline filter for the

EASY-nLC 1000 instrument

6.

From valve B (port 6) to the flow sensor inlet

2

3

1

6

4 5

7.

Valve B

8.

Valve B (port 1) to the pressure sensor

–or– the pressure sensor

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Troubleshooting

Troubleshooting a System Blockage

To troubleshoot a system blockage when only the Test Solvent B test fails

1. To determine if the blockage is in the mixing Tee, do the following: a. Make sure that the Column Out line is connected to the Waste In line with a union.

b. Remove the solvent line from the mixing Tee that connects it to flow sensor B.

c.

Repeat the Back Pressure script with the Test Solvent B check box selected.

d. Depending on the test results, do one of the following:

• If Test solvent B passes, clean the mixing Tee and if necessary replace it.

• If Test solvent B fails, go to step 2 .

2. To determine if the blockage is in the solvent line that connects the B flow path to the mixing Tee, do the following: a. Disconnect the solvent line from the outlet of flow sensor B.

b. Place valve B in position 1–6 as follows: i.

Press Home > Overview.

ii. Press the valve B icon.

iii. In the Valve B dialog box, press 1–6.

c.

Set pump B to deliver 2 μL/min as follows: i.

Press the pump B icon.

ii. In the flow rate box, type 2 μL/min.

iii. Press Start.

iv. Leave the dialog box open.

For more information about the direct controls for the pump, see

“Using the Pump

Controls” on page 205 .

d. Monitor the Pressure readback box for pump B. Then, depending on the back pressure, do one of the following:

• If the pressure is <20 bar, the solvent line is blocked.

– For the EASY-nLC II instrument, replace the solvent line with a new fused silica line. Then, to confirm that the blockage was in the discarded solvent line, repeat the Back Pressure script with the Test Solvent B check box selected.

– For the EASY-nLC 1000 instrument, place valve B in position 1–2 to release the pressure. Remove the blocked nanoViper line and replace it with a new nanoViper line. Then, to confirm that the blockage was in the discarded solvent line, repeat the Back Pressure script with the Test Solvent B check box selected.

• If the pressure is >20 bar, keep pump B running and go to step 3 for the

EASY-nLC 1000 instrument or

step 4 for the EASY-nLC II instrument.

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Troubleshooting

Troubleshooting a System Blockage

3. For the EASY-nLC 1000 instrument, to determine if the inline filter connected to the flow sensor outlet is blocked, do the following: a. Remove the inline filter from the flow sensor outlet.

b. Depending on the back pressure, do one of the following:

• If the pressure is <20 bar, the inline filter is blocked. Replace it with a new filter.

• If the pressure is >20 bar, reconnect the inline filter to flow sensor A, and then go

to step 4 .

4. To determine if flow sensor B is blocked, do the following: a. Disconnect the solvent line from the inlet of flow sensor B, and then monitor the pressure.

b. Depending on the pressure, do one of the following:

• If the pressure is <20 bar, flow sensor B is blocked. Replace it as described in

“Replacing a Flow Sensor” on page 92

.

• If the pressure is >20 bar, keep pump B running and go to step 5 for the

EASY-nLC 1000 instrument or

step 6 for the EASY-nLC II instrument.

5. For the EASY-nLC 1000 instrument, to determine if the inline filter connected to the inlet of flow sensor B is blocked, do the following: a. Remove the inline filter from the flow sensor outlet.

b. Depending on the back pressure, do one of the following:

• If the pressure is <20 bar, the inline filter is blocked. Replace it with a new filter.

• If the pressure is >20 bar, reconnect the inline filter to flow sensor A, and then go

to step 6 .

6. To determine if the line that connects valve B to flow sensor B is blocked, do the following: a. Disconnect the line from port 6 of valve B, and then monitor the pressure.

b. Depending on the pressure, do one of the following:

• If the pressure is below the threshold, the solvent line is blocked. Discontinue this troubleshooting procedure, and replace the solvent line that connects port 6 of valve B to the flow sensor inlet.

Instrument

EASY-nLC II

EASY-nLC 1000

Pressure threshold

20 bar

10 bar

• If the pressure is above this threshold, keep the pump running and go to

step 7 .

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Troubleshooting

Troubleshooting a System Blockage

7. To determine if valve B is blocked, do the following: a. Remove the solvent line from port 1 of valve B, and then monitor the pressure.

b. Depending on the pressure, do one of the following:

• If the pressure is below the threshold, valve B is blocked. Go to

“Maintaining the

Rotary Valves” on page 74 .

Instrument

EASY-nLC II

EASY-nLC 1000

Pressure threshold

20 bar

10 bar

• If the pressure is above the threshold, keep the pump running and go to step 8 .

8. To determine if either the line that connects valve B to pressure sensor B is blocked or pressure sensor B is blocked, do the following: a. Remove the solvent line that connects valve B to pressure sensor B from pressure sensor B, and then monitor the pressure.

b. Depending on the pressure, do one of the following:

• If the pressure is below the threshold, the solvent line is blocked. Replace it.

Instrument

EASY-nLC II

EASY-nLC 1000

Threshold pressure

20 bar

10 bar

• If the pressure is >20 bar, pressure sensor B is blocked. Replace it as described in

“Replacing a Pressure Sensor for the PLU Pump” on page 90

.

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Troubleshooting

Troubleshooting the Autosampler Aspiration and Calibration

Troubleshooting the Autosampler Aspiration and Calibration

Troubleshooting the autosampler aspiration requires these tools and materials:

• 1/4 in. open-ended wrench

• Powder-free safety gloves

• Sample vial filled with solvent A

• Microtiter plate (if you are using a microtiter plate format)

To troubleshoot the autosampler aspiration of sample into the needle

1. Make sure that the W3 wash bottle contains sufficient solvent.

2. Use the Flush Air script to remove air from pump S as follows: a. Press Maintenance > Scripts.

b. In the Category list, select Prepare.

c.

In the Name list, select Flush Air.

d. Press the Parameters tab.

e.

Select the Flush Pump S check box.

f.

For Flush Threshold [μL], enter 10 for the PLF model pump or 12 for the PLU model pump (see

Figure 231 ).

Figure 231. Flush Air script

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Troubleshooting

Troubleshooting the Autosampler Aspiration and Calibration g. Press Start.

The script runs until the flush threshold reaches the specified value or one of these error messages appears on the Output page:

• Aborting unable to build pressure

• Aborting check solvent level

3. Depending on whether the Flush Air script passed or failed, do one of the following:

• If the Flush Air script failed, leave the Maintenance > Scripts page open and go to

step 4 to determine if the autosampler needle is blocked.

• If the Flush Air script passed, leave the Maintenance > Scripts page open and go to

step 5 to determine if the autosampler requires calibration.

4. To determine if the autosampler needle is blocked, do the following: a. Using a 1/4 in. open-ended wrench, disconnect the PEEKsil autosampler needle from port 1 of valve S. b. Disconnect the Teflon waste line from port 3 of valve W and connect it to port 1 of valve S.

c.

Place the free end of the Teflon line in an HPLC vial filled with solvent A.

d. To open the Purge script, select Purge in the Name list. e.

Select the Purge Pump S check box.

f.

Press Start.

g. Verify that pump S aspirates solvent from the HPLC vial.

h. Do one of the following:

• If pump S can aspirate solvent through the Teflon waste tubing, the needle is blocked. Reconnect the Teflon waste tubing to port 3 of valve W and replace the

autosampler needle (see “Replacing the Autosampler Needle” on page 84

).

• If pump S cannot aspirate solvent through the Teflon waste tubing, go to

“Troubleshooting a Pump that Fails the Flush Air Script” on page 212 .

5. To determine if the autosampler needle is going to the correct position and depth, go to

“Checking Sample Pickup” on page 263 .

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Troubleshooting

Checking Sample Pickup

Checking Sample Pickup

When you run the Sample Pickup script, the autosampler withdraws a specified volume from a specified position. To verify the volume withdrawn by the autosampler, you must place a known volume into the specified position, and then measure the remaining volume after you run the script. You can use a pipette or an analytical balance with microgram accuracy to determine the initial and final volumes.

To check sample pickup from a microtiter plate or vial

1. Fill the well or vial with a 12 μL of solvent A.

2. Weigh the microwell plate or vial.

3. Insert the tray into the tray compartment.

4. Press Maintenance > Scripts.

5. Run the Sample Pickup script with a pickup volume of 10 μL and your default flow

(see

Figure 232 ).

Figure 232. Sample Pickup script under the Maintenance menu tab

Thermo Scientific

6. After the EASY-nLC application finishes the script, do the following: a. Eject the tray.

b. Check that 2 μL is left in the well or vial by reweighing the microplate or vial or by checking the volume with a pipette.

c.

If more than 2 μL remains, recalibrate the autosampler, run the Flush Air script, and then rerun the sample pickup check.

For information about calibrating the autosampler, see Chapter 7

. For information about the Flush Air script, see

“Prepare – Flush Air” on page 33 .

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Troubleshooting

Troubleshooting Communication Problems

Troubleshooting Communication Problems

To troubleshoot communication problems, see these topics:

“Network Access” on page 264

“Network Connection Failures” on page 265

“Testing the Network Connection to the EASY-nLC Computer” on page 266

Network Access

Table 25

contains troubleshooting tips for network access problems.

Table 25. Network access problems

Symptom

System is not responding after pressing Save Configuration on the Network page under

Configuration.

Cannot access the EASY-nLC system through the network.

Possible causes

Length of completion time to update the network settings

Changed network address

Action

As indicated, wait while the system changes the network settings.

The stand-alone EASY-nLC instrument is not connected to a laboratory LAN port with intranet access.

The LC/MS system is not connected to a laboratory LAN port with intranet access.

Check the network address of the EASY-nLC system on the Configuration > Network page.

Connect the stand-alone EASY-nLC instrument to a laboratory LAN port with intranet access.

Connect the data system computer for the

LC/MS system to a laboratory LAN port with intranet access.

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Troubleshooting Communication Problems

Network Connection Failures

Table 26

lists the possible causes of a network connection failure. Figure 233

shows the

Thermo EASY-nLC Configuration dialog box that you access from the Thermo Foundation

Instrument Configuration application. When the EASY-nLC instrument cannot establish communication with the data system computer, the Connection Failed message appears when you click Test Connection.

Figure 233. Network connection failure

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Troubleshooting

Troubleshooting Communication Problems

Table 26. Possible causes of a network connection failure

Symptom Possible cause

The connection between the

EASY-nLC instrument and the virtual interface for the Xcalibur data system fails.

The network connection has been unexpectedly disabled.

Action

Test the network connection as described in

“Testing the Network Connection to the

EASY-nLC Computer” on page 266 .

Close down the EASY-nLC instrument as described in

“Closing Down the EASY-nLC

Instrument” on page 21

, and then turn off the power. Restart the Xcalibur data system computer. Turn on the EASY-nLC instrument and log in as an administrator. Retest the connection.

Test the connection. An antivirus program is blocking port 6666.

Make sure that the data system computer is not connected to the Internet, and then uninstall the antivirus program.

The network firewall is blocking port 6666.

Test the connection.

Connection between the data system computer and the

EASY-nLC instrument has been lost.

Remove the firewall.

Test the connection.

Testing the Network Connection to the EASY-nLC Computer

If the connection between the EASY-nLC instrument and the data system computer fails when you click Test Connection in the Thermo EASY-nLC Configuration dialog box of the

Foundation Instrument Configuration window, follow this procedure.

To test the network connection between the data system computer and the EASY-nLC instrument

1. From the Windows Start menu, choose Run.

The Run dialog box opens.

2. In the Open box, type cmd.

3. Press OK.

The Command Prompt window opens.

4. Type telnet IP address for the EASY-nLC system 6666 (see Figure 234

).

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Troubleshooting

Troubleshooting Communication Problems

Figure 234. Command Prompt window with the default IP address for the EASY-nLC instrument

5. Press the ENTER key.

• If the connection is set up correctly, a login prompt appears in the Telnet window

(see

Figure 235 ).

• If the connection is not set up correctly, the following message appears:

Could not open connection to host.

Figure 235. Telnet window with login prompt

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Troubleshooting

Verifying that the LC/MS System Is Properly Grounded

Verifying that the LC/MS System Is Properly Grounded

When you use the EASY-nLC instrument as an inlet to an MS detector, the LC/MS system might require two or more fourplex outlets. The interconnected electrical outlets must have a common point to one ground connector. Connecting the hardware to external grounds at different potentials can create a ground loop that causes noise and interference and can damage the contact closure relays on the instrument’s back panel.

IMPORTANT The EASY-nLC instrument, the MS or LC detector, and the (optional) data system hardware must have a common ground for several reasons:

• Improper grounding or no grounding can result in shock and fire hazards in case of instrument malfunction.

• Connecting the hardware to external grounds at different potentials can create a ground loop that causes noise and interference.

• Improper grounding or no grounding can damage the contact closure relays on the instrument’s back panel. Fixing damaged relays requires the replacement of the instrument’s built-in computer.

CAUTION Electrical safety regulations require that the EASY-nLC instrument be grounded to prevent electrical shock and a fire hazard in case of malfunction.

To verify a common ground between the EASY-nLC instrument and external hardware

1. Connect the EASY-nLC instrument to line power, but do not turn on the instrument.

2. Using a multimeter, measure the resistance between the EASY-nLC chassis and the external hardware chassis (for example, the mass spectrometer chassis or the data system computer) as follows: a. Connect one probe to an unpainted surface on the EASY-nLC chassis and connect the other probe to an unpainted surface on the external hardware chassis

(see

Figure 236 ).

Figure 236. Resistance measurement

Multimeter set up to take a resistance measurement b. Ensure that the resistance measurement is from 0 to 10 ohms.

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Calibrating the Autosampler’s XYZ Robot

To calibrate the autosampler’s XYZ robot, follow the instructions in this chapter.

Contents

Replacing the Autosampler Adapter Plate

Managing Plate Formats

Preparing the Autosampler for the Calibration Routines

Calibrating Plates

Calibrating the Wash Bottle and Extra Vial Positions

For the ASC autosampler, the calibration covers the following areas of the autosampler.

Microtiter plate or vial plate

Extra vials

Waste bottle with needle wash insert

Wash solvent bottles

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Calibrating the Autosampler’s XYZ Robot

Replacing the Autosampler Adapter Plate

For the legacy ASA autosampler, the calibration covers the following areas of the autosampler.

Microtiter/target plate

Wash/waste bottles

Calibrate the autosampler in the following situations:

• When you want to use a new type of vial or microwell plate.

• When you would like to replace the needle.

• For problems with sample pickup; for example, the needle is not placed in the center of the well, or not all of the liquid is picked up from one or more of the vials.

• For problems with the wash/waste bottles; for example, the needle is not placed in the center of the bottle.

Replacing the Autosampler Adapter Plate

The EASY-nLC system ships with the vial adapter plate installed. The microtiter adapter plate comes in the accessory kit. To load microtiter (microwell) plates into the tray compartment, you must replace the vial adapter plate with the microtiter adapter plate.

The vial adapter plate holds 48 vials + 6 extra vials. The microtiter adapter plate holds one

96-well plate or one 384-well plate.

Follow the procedure for the autosampler model in your EASY-nLC system to switch between the two adapter plates as necessary.

“Replacing the Adapter Plate in the ASC Model Autosampler” on page 271

“Replacing the Adapter Plate in the ASA Model Autosampler” on page 272

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Replacing the Autosampler Adapter Plate

Replacing the Adapter Plate in the ASC Model Autosampler

The ASC model is the current autosampler model for both the EASY-nLC II and the

EASY-nLC 1000 systems.

To replace the adapter plate in the ASC model autosampler

1. Press Home > Overview.

The Overview page of the Home menu opens.

2. Make sure that the area in front of the tray compartment is clear, and then press

Eject Plate.

The tray compartment opens. Figure 237

shows the vial adapter plate mounted to the tray holder. The four studs on the bottom side of the plate fit snugly into the mounting holes on the tray holder.

Figure 237. Open tray compartment

3. Remove the current adapter plate by pulling it up and away from the tray holder

(see

Figure 238 ).

Figure 238. Removing the vial adapter plate before installing the microwell plate adapter plate

Mounting stud

Thermo Scientific

4. Install the other adapter plate in the autosampler.

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Replacing the Autosampler Adapter Plate

Replacing the Adapter Plate in the ASA Model Autosampler

The ASA model autosampler is the original autosampler installed in the EASY-nLC II system.

To replace the adapter plate in the ASA autosampler

1. Eject the tray as follows: a. Press Home > Overview.

b. Press Eject Plate.

Figure 239

shows an ejected tray.

Figure 239. Tray compartment of the ASA model autosampler

2. Using the 2.5 mm hex wrench provided with your system, remove the screws that secure the plate to the tray.

The adapter plate is mounted with 4 or 6 hex screws as shown in

Figure 240 .

Figure 240. Screws that secure the adapter plate to the tray

4 hex screws 6 hex screws

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Replacing the Autosampler Adapter Plate

3. Place the new adapter plate on the tray (see

Figure 241 ).

Figure 241. Tray for the ASA model autosampler

4. Fasten the screws in the following balanced order: a. Screw in the upper right corner.

b. Screw in the lower left corner.

c.

Screw in the upper left corner.

d. Screw in the lower right corner.

e.

Screws in the middle.

Note Mounting the screws in a balanced order can avoid distortion of the adapter plate.

The autosampler is now ready for new plate formats. For information about selecting a plate format and creating a new plate format, see

“Managing Plate Formats” on page 274

. After you create a new plate format, perform the calibration routines described in this chapter.

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Managing Plate Formats

Managing Plate Formats

When the EASY-nLC system is initially installed, the application includes plate formats for vials and microtiter plates

Follow the appropriate procedure to use an existing plate format, create your own plate format, or delete a plate format that you no longer need.

“Selecting a Plate Format,”

on this page

“Creating a New Plate Format” on page 275

“Deleting a Plate Format” on page 277

Selecting a Plate Format

To select the plate format that you want to use

1. Open the Tools view for the autosampler as follows: a. Press Maintenance > Devices. b. In the Devices list, select the autosampler device.

c.

Press the Tools tab.

Figure 242

shows the Tools view for the autosampler.

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Figure 242. Tools view for the autosampler

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Managing Plate Formats

Copy To button

Calibrate button

2. Select a plate format from the Plate list.

3. Press Use.

Selected plate format

Tip If you are using a Thermo Scientific software application to control your liquid chromatograph/mass spectrometer (LC/MS) system, the plate selection for the

EASY-nLC system in your Thermo Scientific instrument control software must match the touch-screen application selection.

Creating a New Plate Format

To create a new plate format

1. Open the Tools view for the autosampler as follows: a. Press Maintenance > Devices. b. In the Devices list, select the autosampler device.

c.

Press the Tools tab.

Figure 242

shows the Tools view for the autosampler.

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Managing Plate Formats

2. Do one of the following:

• To create a new plate format, go to

step 3

.

• To copy an existing plate format (copies the format selection), go to step 4 .

3. Create a new plate format as follows: a. Press New.

The Create New Plate Format dialog box opens ( Figure 243

).

Figure 243. Create New Plate Format dialog box b. In the Format list, select a plate format: 6 × 8 (vials), 8 × 12 (96-well plates), or

16 × 24 (384-well plates).

c.

In the Name box, enter a name for the new plate format.

d. Press Accept.

The new plate name appears in the plate list with the text [uncalibrated] appended on the right.

e.

Go to “Calibrating Plates” on page 280 to calibrate the vial or well positions.

4. Copy an existing plate format as follows: a. Select an existing plate format from the Plate list. b. Press Copy To.

The Copy Plate Format dialog box opens.

c.

In the Name box, type a name for the new plate format.

d. Press Accept.

e.

Go to “Calibrating Plates” on page 280 to calibrate the vial or well positions.

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Managing Plate Formats

Deleting a Plate Format

To delete a plate format

1. Open the Tools view for the autosampler on the Devices page of the Maintenance menu

(see

step 1 on page 275

).

2. Select a plate format from the Plate list.

Note You cannot delete the plate in use. When you select the plate in use (highlighted in bold text), the Delete button becomes unavailable.

3. Press Delete.

The Delete Plate Data dialog box opens.

4. Press Accept.

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Preparing the Autosampler for the Calibration Routines

Preparing the Autosampler for the Calibration Routines

To prepare the autosampler for the calibration routines, you must remove the left side panel from the instrument and load the appropriate adapter plate, vials or microtiter plate, wash bottle (W4) with insert, and (optional) extra vial into the tray compartment.

CAUTION Calibrating the autosampler requires removal of the side plate and visual inspection of the autosampler needle holder. Because the autosampler compartment contains moving parts and sharp needles, make sure to keep hands clear when operating the autosampler during calibration.

To prepare the autosampler for the plate calibration routine

1. Install the appropriate adapter plate.

2. To calibrate the vial or well positions on a plate, do one of the following:

• To calibrate the 6 × 8 vial positions, remove the vial caps from two vials and load them into the adapter plate in positions A1 and F8.

• To calibrate a microwell plate, remove the mat or cover from the microwell plate and load the microwell plate onto the adapter plate.

3. Open the Tools view for the autosampler as follows: a. Press Maintenance > Devices.

b. In the Devices list, select the autosampler.

c.

Press the Tools tab.

4. In the Plate area, do the following: a. Select the appropriate plate format from the list, or create a new plate format

(To create and select plate formats, see “Managing Plate Formats” on page 274

).

b. Press Use.

5. Remove the left side panel so that you can watch the movement of the needle.

To calibrate the plate, go to the next topic “Calibrating Plates.”

To prepare the autosampler for the V1 and W4 calibration routine

1. Place an empty vial in position V1.

The V1 position is not available for the microtiter plate formates.

2. Prepare the bottle for position W4 (used for ejecting waste and cleaning the outside of the injection needle).

3. For information about preparing the W4 wash bottle, refer to the EASY-nLC Series

Getting Started Guide.

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Preparing the Autosampler for the Calibration Routines

4. Install the wash bottle in position W4.

IMPORTANT For the autosampler to determine the appropriate depth for the needle, the wash bottle in position W4 must contain the needle wash insert.

5. Remove the left side panel so that you can watch the movement of the needle.

To calibrate the wash bottle and extra vial positions, go to “Calibrating the Wash Bottle and

Extra Vial Positions” on page 285 .

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Calibrating Plates

Calibrating Plates

For information about selecting or creating a plate format, see “Managing Plate Formats” on page 274 .

IMPORTANT When calibrating the XYZ robot for vial or microwell plates, make sure to remove the vial caps or plate covers. Keeping the caps or covers on makes the needle susceptible to bending.

During a sample run, the z-axis needle holder moves to the specified sample position along the xy-plane. When the z-axis needle holder reaches the specified location, the needle descends along the z-axis into the sample vial or well.

Calibrating plates requires determining the xyz positions for the top left and bottom right vial

or well. Table 27

lists the top left and bottom right positions for the three plate formats.

Table 27. Top left and bottom right positions on the plate formats

Plate format

6 × 8

8 × 12

16 × 24

Top left position

A1

A1

A1

Bottom right position

F8

H12

P24

To calibrate plates

1. If you have not already done so, prepare the autosampler for the plate calibration routine

(see

“To prepare the autosampler for the plate calibration routine” on page 278

).

2. Open the Autosampler Plate Calibration dialog box as follows: a. Press Maintenance > Devices.

b. In the Devices list, select the autosampler.

c.

Press the Tools tab.

The Tools view opens (see Figure 244

).

Figure 244. Tools view for the autosampler

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Calibrating Plates d. In the Plate area of the Tools view for the autosampler, press Calibrate

(see

Figure 244 on page 280

).

The Autosampler Plate Calibration dialog box opens.

Figure 245

shows the dialog box at the beginning of the plate calibration routine.

The top left and bottom right positions are highlighted in green. The position readbacks for the xyz axes are unpopulated, .

Figure 245. Plate view in the Autosampler Plate Calibration dialog box

(3)

(1)

(5)

(2)

(4)

3. Press the green vial or well in the upper left corner of the plate view (1).

The needle moves to the current saved coordinate. The saved coordinates for the x and

y axis appear on the left side of the slash, the needle does not descend into the vial or well, and the z-axis value on the left side of the slash remains at 0.

The values on the left side of the slash change as you adjust the needle position.

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Calibrating Plates

4. Center the needle above the vial or well as follows:

• Press the Needle In/Out (3) buttons to adjust the x-axis position.

• Press the Needle Left/Right (4) buttons to adjust the y-axis position.

y axis

1

In

A

x axis

Out

Right

Left

5. Using the step buttons in the Needle Up/Down area (5), adjust the needle height as follows: a. Using the big step button (), lower the needle until it reaches a level of 2–3 mm

(0.08–0.12 in.) above the vial or well. b. Using the small step button (), slowly lower the needle until it reaches the bottom of the vial or well.

Tip For the ASC autosampler, the O-ring at the top of the needle holder rises a

little when the needle reaches the bottom of the vial or well (see Figure 246

).

Figure 246. O-ring pushed slightly above the ASC needle holder

Observe when the O-ring moves up a bit, or gently hold needle tubing as you press the step button.

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Calibrating Plates

Tip For the ASA autosampler, do one of the following to determine the bottom position of the vial or well:

• To feel when the needle reaches the bottom of the vial or well, hold the needle gently as you press the down arrow on the touch-screen interface.

• To see when the needle reaches the bottom of the vial or well, draw a mark on the needle and observe the needle mark as you press the down arrow on the touch-screen interface. When the needle mark moves up a bit, the needle has reached the bottom of the vial.

Figure 247 shows the ASA needle holder and

needle.

Figure 247. ASA autosampler needle holder

Use a black marker pen to make a line on top of the

PEEKsil tubing.

c.

Press the small step button () once to raise the needle slightly off the bottom of the vial or well.

The appropriate needle height is one small step higher than the bottom of the vial or well.

6. When you are satisfied with the current xyz-coordinate for the vial or well, press Capture.

The color of the vial or well on the plate graphic changes from green to red and the position readbacks for the xyz axes are unpopulated,

(see Figure 248

).

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Calibrating Plates

Figure 248. Plate calibration captured for the A1 position

7. Press the green vial or well in the lower right corner of the plate view.

The needle moves to the current saved coordinate. The saved coordinates for the x and

y axis appear on the left side of the slash, the needle does not descend into the vial or well, and the z-axis value on the left side of the slash remains at 0.

8. Repeat

step 4

through

step 6

on that vial or well.

9. Press Save and then press Close to close the calibration dialog box.

10. Replace vial caps and plate covers as necessary.

11. Go to the next procedure to calibrate the W4 bottle position or reinstall the EASY-nLC instrument’s left side panel.

IMPORTANT To maintain a stable temperature inside the tray compartment, the left side panel must be installed.

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Calibrating the Wash Bottle and Extra Vial Positions

Calibrating the Wash Bottle and Extra Vial Positions

In addition to holding the sample trays, the EASY-nLC tray compartment holds four wash bottles (W1–W4) and six extra vials (V1–V6).

To calibrate the W4 wash bottle position, the extra vial positions, or both

1. If you have not already done so, prepare the autosampler for the extra vial and wash bottle calibration as described in

“To prepare the autosampler for the V1 and W4 calibration routine” on page 278

.

2. Open the Autosampler Vials/Wash Calibration dialog box as follows: a. Press Maintenance > Devices.

b. In the Devices list, select the autosampler.

c.

Press the Tools tab below the Devices list.

d. In the Vials/Wash area, press Calibrate.

The Autosampler Vials/Wash Calibration dialog box opens (see Figure 249

).

3. Press the bottle container in W4 (1) (see

Figure 249

).

The needle moves to the current saved coordinate. The saved coordinates for the x and

y axis appear on the left side of the slash, the needle does not descend into the bottle, and the z-axis value on the left side of the slash remains at 0.

Figure 249. Vials/Wash view in the Autosampler Vials/Wash Calibration dialog box

(2)

(5)

(4)

(1)

(3)

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Calibrating the Wash Bottle and Extra Vial Positions

4. Center the needle above the bottle as follows:

• Press the Needle In/Out (2) buttons to adjust the x-axis position.

• Press the Needle Left/Right (3) buttons to adjust the y-axis position.

y axis

In

x axis

Out

Right

Left

5. Using the step buttons in the Needle Up/Down area (5), adjust the needle height as follows: a. Using the big step button (), lower the needle until it reaches a level of 2–3 mm

(0.08–0.12 in.) above the bottle. b. Using the small step button (), slowly let the needle enter the hole and lower the needle until it reaches the bottom of the wash insert in the bottle.

Note The O-ring at the top of the needle holder rises a little when the needle reaches the bottom of the well.

c.

Press the large step button () twice to raise the needle off the bottom of the wash bottle insert.

Two large steps higher than the bottom of the wash bottle insert is the appropriate needle height.

Note The ASA autosampler has a limit of 850 steps on the z axis (vertical). This limit is set to avoid bent needles. You might be able to go for more than 850 steps during calibration, but the needle does not go more than 850 steps during normal work.

6. Press Capture when you are satisfied with the current xyz-coordinate for position W4.

The color of the bottle container number changes from green to red.

7. (Optional) To calibrate the extra vial positions for the 6 × 8 plate format, do the following: a. Press V1 vial (5) to select it.

b. Repeat

step 4

through

step 6

for vial V1.

8. Press Save and then press Close to close the calibration dialog box.

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Remote Support

The EASY-nLC instrument includes a remote support feature that enables Thermo Fisher

Scientific Technical Support to diagnose and troubleshoot your EASY-nLC system remotely while the instrument is connected to the support server. When you connect the instrument to the remote server, you can also install the latest firmware file for the touch-screen application.

Technical Support cannot initiate the communication link between your

EASY-nLC instrument and the support server. You must initiate this communication link from your EASY-nLC instrument.

To connect the EASY-nLC instrument to the support server or download system files to a removable storage device, follow these procedures.

Contents

Connecting the EASY-nLC Instrument to the Support Server

Saving System Files on a USB Removable Storage Device

Downloading the Latest Firmware File

IMPORTANT To establish communication with the support server, you must first connect the EASY-nLC instrument directly to your local network, and then open the communication link for remote support through the touch-screen software. Thermo

Fisher Scientific Technical Support cannot initiate this communication link.

The EASY-nLC instrument uses Secure Shell (SSH™), a network protocol for secure data communication, to connect to the support server, which uses the Linux™ operating system.

To use the remote support feature, the firewalls for your local network must allow outgoing TCP/IP traffic from the EASY-nLC system to the support server at

IP address 195.41.108.93 port 22.

When you establish the communication link with the support server, the support server triggers the file compression utility on the EASY-nLC instrument to build a zip file with the appropriate log files, and then retrieves the zip file from the EASY-nLC instrument.

Meanwhile, the EASY-nLC instrument sends an e-mail with your supplied message and contact information to Thermo Fisher Scientific Technical Support.

After you send a message through the remote support feature, a member of the Thermo

Fisher Scientific Technical Support team will contact you.

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Connecting the EASY-nLC Instrument to the Support Server

Connecting the EASY-nLC Instrument to the Support Server

Use this procedure to establish a communication link between the EASY-nLC instrument’s computer and the support server.

To open a communication link between the EASY-nLC system and the support server

1. Using a shielded Ethernet cable, connect the LAN port on the back panel of the

EASY-nLC system (see

Figure 250 ) directly to a laboratory LAN port that provides access

to the Internet.

To use the remote support feature, the firewalls for your local network must allow outgoing TCP/IP traffic from the EASY-nLC system to the support server at

IP address 195.41.108.93 port 22.

Figure 250. Ethernet connection between the EASY-nLC system and the laboratory LAN port

USB LAN

LAN MAC ADDRESS: 00-6--E0-45-4E-28

RS-232

HARD

DRIVE

MONITOR

P-BUS

IN3 OUT3 IN2 OUT2 IN1 OUT1

I

O

110-120V▼

220-240▼

120/230 V : 50/60 Hz: 250 W

Fuse at 120 V– : T5 AL 250 V

Fuse at 230 V– : T2.5 AL 250 V

LAN port with

Internet capability

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Connecting the EASY-nLC Instrument to the Support Server

2. If you have not already set up the support server configuration, set it up as follows: a. Press Configuration > Network.

The Network page of the Configuration menu opens (

Figure 251 ).

Figure 251. Network page of the Configuration menu

Thermo Scientific b. In the LAN area, do the following: i.

Select Internet in the Profile list.

ii. Press Apply.

c.

In the Support Server area, do the following: i.

Enter the following IP address: 195.41.108.93.

ii. Press Apply.

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Connecting the EASY-nLC Instrument to the Support Server

3. Send a support request message and a zip file containing the appropriate log files to

Technical Support as follows: a. Press Maintenance > Support.

The Support page of the Maintenance menu appears (see

Figure 252 ).

Figure 252. Support page of the Maintenance menu b. In the Connect to EASY-nLC Support area, press Connect.

Depending on whether the instrument has the latest firmware file installed, one of these dialog boxes appears:

• If the instrument has the latest firmware file installed, the Message to Support

Team dialog box appears (see

Figure 253 ).

• If the instrument does not have the latest firmware file installed, the Upgrade is

Available dialog box appears (see

Figure 256 on page 296

).

c.

If the Upgrade is Available dialog box appears, do one of the following:

• Upgrade the software as described in “Downloading the Latest Firmware File” on page 296 .

–or–

• Click Skip.

The Message to Support Team dialog box appears.

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Connecting the EASY-nLC Instrument to the Support Server d. Select a support region, provide valid contact information, and describe the problem that you are experiencing.

Figure 253. Message to Support Team dialog box

Thermo Scientific

Note When you place the cursor in the Phone Number box, the program prompts you for a country code and phone number.

Tip When you place the cursor in the Email Address box, the program prompts you to enter an address in this format: name@company.com.

To enter the @ symbol, press Shift. The numbers row changes to the symbols row. Then press @.

e.

Press Accept.

After you enter the requested information, the Status box on the right side of the

Support page should change from Not Connected to Connected.

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Connecting the EASY-nLC Instrument to the Support Server f.

Depending on the connection status, do one of the following:

• If the status changes from Not Connected to Connected, go to step 3g

.

• If the connection fails, see “To troubleshoot a failed connection to the support server” on page 292 .

When the connection fails, the SSH Connection Error dialog box might appear with this message, “Not able to determine connection status. See message.log for details.”

The message.log file is in the admin\systemLog folder on the EASY-nLC system. For information about accessing the admin folder from another computer on your local network, refer to the EASY-nLC Series Getting Started

Guide.

While the message.log file might provide useful information to a member of

Thermo Scientific Technical Support, users should follow the suggested

troubleshooting procedure, “To troubleshoot a failed connection to the support server” on page 292 .

g. After the status changes to Connected, allow enough time for the EASY-nLC system to create the zip file in the admin folder and for the support server to retrieve the zip

file. Then go to step 4 .

After you send a message with the appropriate contact information through the remote support feature, a member of the Thermo Fisher Scientific Technical Support team will contact you.

4. To close the communication link, press Disconnect.

5. To return the system to normal operation, do the following: a. Reconnect the Ethernet cables as appropriate.

For information about connecting the EASY-nLC system to a data system computer, refer to the EASY-nLC Series Getting Started Guide.

b. In the LAN area of the Configuration > Network page, do the following: i.

Select LAN in the Profile list.

ii. Press Apply.

To troubleshoot a failed connection to the support server

1. Check the support server configuration (see step 2 on page 289 ).

2. Verify that the Ethernet cable is securely connected to both the LAN port on the back panel of the EASY-nLC system and the laboratory LAN port.

3. Verify that the Ethernet cable is working.

4. Ask your IT administrator if the network firewalls allow outgoing traffic to port 22 through an SSH connection.

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Saving System Files on a USB Removable Storage Device

Saving System Files on a USB Removable Storage Device

The EASY-nLC system includes network capability. Through the network you can back up your system, export and import batches and methods, analyze your system by examining the log files, and perform many other tasks. However, when you are not connected to the network or the network is broken, or you do not have a computer with network access that is near the

EASY-nLC system, you can use the copy function to copy all the system files on the system to a USB removable storage device, such as a USB flash drive or memory stick.

The EASY-nLC system ships with a USB flash drive for your convenience. But you can use any USB storage device formatted with the file system FAT/FAT16 and at least 128 MB of free space.

To copy system files to a USB storage device

1. Press Maintenance > Support.

The Support page of the Maintenance menu opens (see Figure 252 on page 290 ).

2. Insert the USB storage device into the connector panel on the back of the EASY-nLC instrument. Choose between one of two USB ports on the upper left side of the panel

(see

Figure 254 ).

Figure 254. USB port location on the back panel of the EASY-nLC instrument

Thermo Scientific

3. In the Log File Copy area, select the To USB Removable Storage option.

Note When you select the To Home Directory option, the EASY-nLC system creates zip files in the admin folder.

For information about accessing the admin folder from another computer on your local network, refer to the EASY-nLC Series Getting Started Guide.

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4. Press Copy Log Files.

When the copying is finished, the following confirmation appears.

5. Press Close and remove the USB storage device from the connector panel.

6. Insert the USB storage device into your computer.

Five zipped files appear on the USB storage device (see

Figure 255 and

Table 28 on page 295 ).

Figure 255. Windows XP view of the files on the USB memory stick

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Saving System Files on a USB Removable Storage Device

Table 28. Zipped files created by pressing Copy Log Files

File name

batchlog.zip

etc.zip

export.zip

logfiles.zip

maintlog.zip

Description

Contains one folder for each batch you have run with information on samples, methods, and EASY-nLC device performance.

Contains system configuration files.

Contains exported batches and methods per user and system backup.

Contains system log files with information on overall system performance.

Contains one folder for each maintenance script you have run with information on EASY-nLC device performance.

If you have problems with your system, forward the files to Thermo Fisher Scientific

Technical Support for assistance.

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Downloading the Latest Firmware File

Downloading the Latest Firmware File

If the EASY-nLC instrument does not have the latest version of the touch-screen application, connect the instrument to the remote server and install the appropriate firmware files as described in the following procedure.

To install the latest firmware file

1. If the instrument is not already connected to the remote server, connect the instrument as follows: a. From the touch screen, press Maintenance > Support.

The Support page of the Maintenance menu appears. b. In the Connect to EASY-nLC Support area, press Connect.

If the EASY-nLC instrument does not have the latest version of the touch-screen application, the Upgrade is Available dialog box appears.

Figure 256

shows the message for an instrument with firmware version 2.4.

Figure 256. Firmware upgrade message

2. Click Download.

When the file transfer is complete, the Transfer of Log and Firmware Files Completed dialog box appears.

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Remote Support

Downloading the Latest Firmware File

Figure 257

shows the transfer information for an instrument with firmware version 2.4.

Figure 257. Information for upgrading from a previous version

Thermo Scientific

3. Click OK.

4. To begin the firmware upgrade, do the following: a. Press Exit in the lower-left corner of the touch screen.

A confirmation dialog box opens (see

Figure 22 on page 21 ).

b. Press Power Down.

The EASY-nLC application displays a white screen with a small progress bar. When the progress bar is completely filled, a message appears indicating you can safely turn off the instrument.

c.

Turn off the instrument, and then turn it back on.

d. When the Upgrade button appears, double-tap it.

5. If the complete upgrade requires more firmware files, reconnect to the server and install

the upgrade files as described in step 1

through

step 4

of this procedure.

Table 29

lists the software versions that you must install to upgrade the software to the current version.

Table 29. Software installation matrix

Current version

2.4 #2923

2.5 #3032

2.7.8.1

2.8.11.1

3.1.4.1

Install versions

2.5 #3032, 2.7.8.1, 2.8.11.1, and 3.1.4.1

2.7.8.1, 2.8.11.1, and 3.1.4.1

2.8.11.1 and 3.1.4.1

3.1.4.1

current as of January 2013

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Returning the EASY-nLC Instrument

If it is necessary to return the EASY-nLC system to the factory, follow the instructions in this chapter.

Contents

Transport Instructions

Declaration of Contamination

Transport Instructions

To prepare the instrument for transport

1. Press Home > Overview.

2. Press Eject Plate.

3. Remove the plates/tubes.

4. Remove the waste beaker and the wash/waste bottles.

5. Press Insert Plate.

6. Close down the system (see

“Closing Down the EASY-nLC Instrument” on page 21

).

7. Remove the cables on the back side.

Tip For the original ASA autosampler only: Secure the autosampler by using the

transport collars (see Figure 258

) before shipping the instrument. (This is not necessary on the latest ASC model.)

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Returning the EASY-nLC Instrument

Transport Instructions

Figure 258. (ASA autosampler only) Transport collar location

Insert two transport collars after closing down the instrument.

8. Fill out the Declaration of Contamination of Equipment in

“Declaration of

Contamination” on page 301

.

9. Place the EASY-nLC instrument in the original shipping container.

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Returning the EASY-nLC Instrument

Declaration of Contamination

Declaration of Contamination

To return, service, or repair the EASY-nLC instrument, you must complete and submit a

“Declaration of Contamination of Equipment” to Thermo Fisher Scientific with the returned equipment. Qualified personnel must review the declaration. Contact Thermo Fisher

Scientific for additional copies of this form or if you have any questions regarding the contents of this declaration.

Description of equipment

Equipment type/model: ______________________ Serial No.: __________________

Date of receiving/purchasing equipment: _________________________

Reason for return: [ ] Maintenance [ ] Repair [ ] End of test

[ ] ______________________________________

Describe symptoms and problems: ___________________________________________

______________________________________________________________________

Equipment condition

Has the equipment been used?

[ ] Yes [ ] No

Has the equipment been exposed to potentially harmful substances?

[ ] Yes [ ] No

If yes, attach list of all known harmful substances including chemical name and symbol, and precautions associated with the substances.

Were any of the harmful substances

• Radioactive?

• Toxic?

• Corrosive?

[ ] Yes

[ ] Yes

[ ] Yes

[ ] No

[ ] No

[ ] No

• Explosive?

[ ] Yes [ ] No

Has the equipment been properly decontaminated and/or cleaned before being returned?

[ ] Yes [ ] No

Legally binding declaration

I hereby declare that the information supplied on this form is accurate and sufficient to judge any contamination level.

Name: ________________________________________________________________

Job title: ____________________ Organization: _____________________________

Address: _______________________________________________________________

Telephone: ________________ Fax: ________________ E-mail: __________________

Signature: ______________________________________________________________

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Error Codes

Table 30

lists the possible error codes.

Table 30. Error codes (Sheet 1 of 8)

No.

Issue code name

1

2

ERROR_IO_CLOSING_

FILE

ERROR_IO_WRITING_

TO_FILE

3

Description

Failed closing log file

IOException

4

5

6

ERROR_IO_NO_FILE_

OPEN

ERROR_IO_CREATE_

FILE

ERROR_IO_CREATE_

FOLDER

ERROR_IO_FILE_

EXISTS

Cause

File system error

I/O error occurred

Log was closed trying to log the following message:

Filesystem error

I/O error occurred Failed creating log file

'LOG_FILE_NAME'

Failed creating log folder

'LOG_FOLDER_NAM

E'

Failed creating log

'LOG_FILE_NAME

AND_PATH' (file already exists)

I/O error occurred

Filesystem error

7

8

9

ERROR_IO_FILE_CAN

NOT_WRITE

ERROR_BATCH_BEIN

G_EDITED

ERROR_DEVICE_

ACCESS_FAILED

Failed creating log

''LOG_FILE_NAME_

AND_PATH' (file not writable)

It appears the current batch is being edited, please close or reschedule it.

Failed accessing device

DEVICE_ID

Filesystem error

[see issue description]

Missing the hardware component driver

Solution

Internal filesystem error -

Replace the hard drive.

Internal filesystem error -

Replace the hard drive.

Internal filesystem error -

Replace hard drive

Internal filesystem error -

Replace the hard drive.

Internal filesystem error -

Replace the hard drive.

Internal filesystem error -

Replace the hard drive.

Internal filesystem error -

Replace the hard drive.

Close or reschedule the batch.

Add the device to the list on the Maintenance

> Devices page.

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Error Codes

Table 30. Error codes (Sheet 2 of 8)

No.

Issue code name

10 ERROR_PREP_

GRADIENT_

PRESSURE_BUILD_

FAILED

11 ERROR_PUMP_

STOPPED_EMPTY

Description

Pump B unable to reach 90% A pressure within 30 μL.

Please check for air/leaks!

Cause

Pump unable to reach set pressure during prep gradient stopped (out of solvent).

Out of solvent stopped unexpectedly

(unknown cause).

Solution

Check that solvent bottle is not empty and check for air and leaks in the system.

Refill the solvent reservoir bottle. Check for air and leaks in the system.

Pump hardware issue Check the pump functionality.

12 ERROR_PUMP_

STOPPED_

UNEXPECTEDLY

26 ERROR_QUEUE_

ELEMENT_BEING_

EDITED

27 ERROR_PBUS_IN_

SERVICE_MODE

Current queue element is being edited; stopping.

Trying to execute a job currently being edited

Close or reschedule the batch.

28

29

30

31

32

ERROR_PBUS_

INTERFACE_INIT

ERROR_PBUS_

SEND_BYTES

ERROR_PBUS_

READ_BYTES

ERROR_SENSIRION_

FACTOR_PARSING

ERROR_SENSIRION_

FACTOR_NOT_

FOUND

P-BUS interface in service mode.

Initialization of P-BUS interface failed

Trying to initialize the

RS485 interface while serial port is in the service mode

Check that the RS485 switch on PC is in the

ON position.

[see issue description] Troubleshoot P-BUS communication -

Check that the cables are properly connected.

Error writing to interface

COMM_PORT

[see issue description] Troubleshoot P-BUS communication -

Check that the cables are properly connected.

Error reading from interface COMM_PORT

[see issue description] Troubleshoot P-BUS communication -

Check that the cables are properly connected.

Cycle EASY nLC power.

Failed parsing flow sensor factor from

INFO_COMMAND

Could not parse some string to a double

Flow sensor factor not found in info output.

Factor unknown.

Error getting flow sensor scaling factor

Cycle EASY nLC power.

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Error Codes

Table 30. Error codes (Sheet 3 of 8)

No.

Issue code name

33 ERROR_

SENSIRION_

INIT

34

35

36

37

ERROR_SENSIRION_

NO_DATA

ERROR_SENSIRION_

IN_MEASURE_MODE

ERROR_SENSIRION_

IO

ERROR_SENSIRION_

TIMEOUT

Description Cause Solution

Port COM_PORT does not support reading/Error getting input stream from flow sensor port

COM_PORT

No data received from flow sensor for X seconds

Error getting input from flow sensor

Flow sensor in measurement mode. Stop measurements before sending further commands

Cycle EASY nLC power.

[see issue description] Cycle EASY nLC power /

Check the flow sensor functionality.

Cycle EASY nLC power.

Trying to start a flow sensor that is already started

IO Error while communicating with flow sensor

[see issue description] Check the flow sensor communication/cable.

No response from flow sensor (timeout)

[see issue description] Cycle EASY nLC power -

If the problem persists, replace the flow sensor.

38

39

40

ERROR_SENSIRION_

ECHO_MISMATCH

ERROR_SENSIRION_

BAD_RESPONSE

ERROR_SENSIRION_

IN_IDLE_MODE

Flow sensor echo

"ECHO" does not match command

"COMMAND"

Flow sensor fails to "talk back"

Flow sensor error:

ERROR_MESSAGE/Flo w sensor did not respond with OK, but did not report an error either

See “Replacing a Flow

Sensor” on page 92 .

Rare error - Connect the

EASY-nLC system to remote support.

See “Connecting the

EASY-nLC Instrument to the Support Server” on page 288 .

Flow sensor failing, but does not report it

Rare error - Connect the

EASY-nLC system to remote support.

Cannot calibrate sensor in idle mode

Flow sensor in idle mode during calibration

Cycle EASY nLC power.

Check the flow sensor functionality.

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Error Codes

Table 30. Error codes (Sheet 4 of 8)

No.

Issue code name

41 ERROR_SENSIRION_

CALIBRATION

Description

Cannot store flows larger than 9999 nL/min

Cause

Abnormal value trying to be written to the flow sensor

Access to interface failed

Error occurred when sending COMMAND

Solution

Cycle EASY nLC power.

Check flow sensor functionality

Cycle EASY nLC power.

42 ERROR_RS232_INIT Initialization of interface

"PORT_NAME" failed

54 ERROR_RS485_

COMMAND_VERIFY

COMPONENT_ID

COMMAND failed:

ERROR_MESSAGE

55 ERROR_

MANIPULATOR_

UNKNOWN_RACK_

SHELF

Unknown rack shelf

RACK_SHELF_VALUE

56 ERROR_

MANIPULATOR_

UNKNOWN_RACK_

POS_ID

57 ERROR_

MANIPULATOR_SET_

GRID_CONFIG L

Unknown rack position id

RACK_POSITION_ID_

VALUE

Grid configuration change failed

(AUTOSAMPLER_

RETURN_CODE)

58

59

ERROR_

MANIPULATOR_SET_

WASH_CONFIG

ERROR_

MANIPULATOR_SET_

VIAL_CONFIG

Unknown rack shelf position supplied to driver

Could not move to bad rack position

Troubleshoot P-BUS communication/hardware.

Verify that the sample position is correct.

Verify that the sample position is correct.

Error uploading autosampler plate calibration

(0 = asrOK,

1 = asrBusy,

2 = asrBadLocation,

3 = asrBadCoordinate,

4 = asrEjected,

5 = asrAxisError,

6 = asrChecksumError)

Calibrated position not recognized by software. See

Chapter 7, “Calibrating the

Autosampler’s XYZ Robot.”

Wash configuration change failed

(AUTOSAMPLER_

RETURN_CODE)

Error while saving autosampler wash calibration (see code 57 for asr codes)

Calibrated position not recognized by software. See

Chapter 7, “Calibrating the

Autosampler’s XYZ Robot.”

Vial configuration change failed

(AUTOSAMPLER_RET

URN_CODE)

Error while saving autosampler vial calibration (see code 57 for asr codes)

Calibrated position not recognized by software. See

Chapter 7, “Calibrating the

Autosampler’s XYZ Robot.”

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Error Codes

Table 30. Error codes (Sheet 5 of 8)

No.

Issue code name

60 ERROR_

MANIPULATOR_TRAY

_EJECT

Description

tray eject failed

(AUTOSAMPLER_RET

URN_CODE)

Cause

see code 57 for asr codes

61 ERROR_

MANIPULATOR_TRAY

_LOAD

62 ERROR_

MANIPULATOR_GRID

_ADDRESS

63 ERROR_

MANIPULATOR_

WASH_ADDRESS

64 ERROR_

MANIPULATOR_VIAL_

ADDRESS

65 ERROR_

MANIPULATOR_

NEEDLE_DOWN

66 ERROR_

MANIPULATOR_

NEEDLE_UP tray load failed

(AUTOSAMPLER_RET

URN_CODE) grid address failed

(AUTOSAMPLER_RET

URN_CODE) wash address failed

(AUTOSAMPLER_RET

URN_CODE) vial address failed

(AUTOSAMPLER_RET

URN_CODE) needle down failed

(AUTOSAMPLER_RET

URN_CODE) needle up failed

(AUTOSAMPLER_

RETURN_CODE) see code 57 for asr codes

Could not move to sample position (see code 57 for asr codes)

(see above)

Could not move to wash position (see code

57 for asr codes)

(see above)

Could not move to vial position (see code 57 for asr codes)

(see above)

Could not move needle down (see code 57 for asr codes)

(see above)

Could not move needle up (see code 57 for asr codes)

(see above)

Solution

Verify that the correct plate is configured in the

EASY-nLC software and the Xcalibur VI. Check that no objects in the tray compartment are obstructing the XYZ robot movement. Reset manipulator and run the

Torque script for the ASC model autosampler.

(see above)

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Error Codes

Table 30. Error codes (Sheet 6 of 8)

No.

Issue code name

69 ERROR_VALVE_SET_

POSITION

Description

COMPONENT_ID setPosition failed:

Unrecognized position

POSITION/

COMPONENT_ID setPosition failed:

Problems switching to position POSITION/

COMPONENT_ID needed ATTEMPTS attempts switching to position POSITION

Cause

Valve unable to switch position

Solution

Clean the rotor seal and the stator. Replace the rotor seal if necessary.

If the problem persists, do the following:

• For valve serial number

< V-009800, run the

Valve Tune script.

• For valve serial number

> V-009800, replace the valve.

70 ERROR_PARALLEL_

PORT_WRITE

71 ERROR_PARALLEL_

PORT_NOT_

INITIALIZED

72 ERROR_PARALLEL_

PORT_MASS_SPEC_

BUSY

73 ERROR_PARALLEL_

PORT_MASS_SPEC_

TIMEOUT

Contact closure port

IO error

Contact closure port

IO error

Contact closure port

IO error

Contact closure port

IO error

Contact Thermo Fisher

Scientific.

Contact Thermo Fisher

Scientific.

Contact Thermo Fisher

Scientific.

Contact Thermo Fisher

Scientific.

74 ERROR_PARALLEL_

PORT_INIT

75 ERROR_PARALLEL_

PORT_WRONG_PORT

77 ERROR_DEVICE_LOW

_24_VOLT_SUPPLY

78 ERROR_

MANIPULATOR_INIT

DEVICE: Low voltage

(VALUE V)

79 ERROR_DEVICE_SELF

_TEST

Contact closure port

IO error

Contact closure port

IO error

Contact Thermo Fisher

Scientific.

Contact Thermo Fisher

Scientific.

[see issue description] Faulty 24V supply -

Connect to remote support

DEVICE: Init failed (not ready within 1 minute)

DEVICE: Status bit(s) set after self test:

0xBITS_IN_HEX

[see issue description] Autosampler hardware issue. Cycle EASY-nLC power. If the problem persists, check the P-bus cables to the autosampler.

Error returned from hardware component firmware

Connect the system to the remote support server (see

Chapter 8

).

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Error Codes

Table 30. Error codes (Sheet 7 of 8)

No.

Issue code name

80 ERROR_DEVICE_IN_

BOOT_MODE

81 ERROR_PELTIER_

SPURIOUS_

TEMPERATURE

82 ERROR_DEVICE_LOW

_CURRENT

83

84 ERROR_APPLICATION

_STATE

85 IC_STATUS_FLAG_

WARNING

86

87

88

ERROR_VALVE_

SPURIOUS_ANGLE

IC_STATUS_FLAG

_SEVERE

IC_LC_CONDITION_

UNSATISFIED

IC_STATUS_FLAG_

ERROR

Description Cause

DEVICE: Device in boot mode after device reset

Error returned from hardware component firmware because a

PCB is damaged.

Solution

Contact Thermo Fisher

Scientific.

DEVICE: Spurious temperature

(TEMPERATURE_IN_

DEGREES_CELSIUS)

Cooler sensor defective Replace the cooler.

See “Replacing the

Autosampler Cooler” on page 137 .

Replace the cooler.

DEVICE: Low current consumption

(CURRENT_VALUE A)

DEVICE: Spurious angle in position POSITION

(ANGLE)

HISTORY_TRACE

Peltier element defective

[see issue description]

Application state empty or illegal

For valve serial number

< V-009800, run the Valve

Tune script.

Cycle EASY nLC power.

Act according to the error message.

Waiting for some condition during run unsuccessful.

Monitor reported flag(s) raised (log file only).

Monitor reported flag(s) raised (in user interface, current run stopped).

Failed reaching a specific flow, pressure, etc. within a specified volume/period of time.

Act according to the error message.

Troubleshoot the LC system for leaks and air.

Monitor reported flag(s) raised (in user interface).

See “Troubleshooting the

Results of the System Leak

Test” on page 221 .

Act according to the error message.

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Error Codes

Table 30. Error codes (Sheet 8 of 8)

No.

Issue code name

89 IC_VALVE_MULTIPLE_

ATTEMPTS

90

92

93

IC_DEVICE_

UNSUPPORTED_

OPERATION

IC_BUSINESSLOGIC_

REFILL_DURING_

GRADIENT

IC_SCRIPT_AUTO_

ABORT

Description

Needed 2 attempts to switch to position 1-6

Gradient interrupted by pump refill warning

Maintenance script self-abort

Cause

A switching valve needed multiple attempts to reach the target position.

Solution

Clean the rotor seal and the stator. Replace the rotor seal if necessary.

Probably because a pump ran out of solvent while building/maintaining pressure

If the problem persists, do the following:

• For valve serial number

< V-009800, run the

Valve Tune script.

• For valve serial number

> V-009800, replace the valve.

A device type operation was attempted on a device instance that doesn't support it.

Contact Thermo Fisher

Scientific.

Pump solvent volume too small for next gradient step.

No action is necessary.

This is just a warning that the pumps were refilled during gradient processing.

Check for air and leaks in the subsystem.

See “Troubleshooting the

Results of the System Leak

Test” on page 221 .

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B

Mobile Phase Viscosity

Table 31

lists the viscosities for two-solvent mobile phases consisting of a water/methanol mixture or a water/acetonitrile mixture.

Table 31. Mobile-phase viscosity at 25

C for reversed-phase gradients

Mobile phase (%v organic/water)

25

(cP)

Methanol Acetonitrile

0

10

20

0.89

1.18

1.40

0.89

1.01

0.98

30

40

50

60

1.56

1.62

1.62

1.54

0.98

0.89

0.82

0.72

70

80

90

100

1.36

1.12

0.84

0.56

0.59

0.52

0.46

0.35

Reference: D.P. Herman, A.H. Billiet, and L. de Galan, Journal of Chromatography,

463 (1989) 1

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Consumables and Replacement Parts

To order consumables and replacement parts for the EASY-nLC system, see the ordering information and solvent system schematics in this chapter.

Contents

Ordering Information

Solvent System Schematic for the EASY-nLC II Instrument

Solvent System Schematic for the EASY-nLC 1000 Instrument

Common Replacement Parts

Ordering Information

To obtain ordering information for the EASY-nLC instrument’s spare parts and accessories, go to the following Web sites:

• For the EASY-nLC 1000 instrument, go to: http://www.proxeon.com/productrange/nano_lC_easy-nlc_1000/accessories_spares

• For the EASY-nLC II instrument, go to: http://www.proxeon.com/productrange/nano_lC/accessories_spares

These EASY-nLC parts are consumable parts and not covered by the normal first year Limited

Warranty offered by Thermo Fisher Scientific, or any other service contract agreement containing extended warranty coverage.

• All flow lines including Tee-pieces and fittings (nuts, ferrules, sleeves, and valve stops)

• Check valves

• Solvent filters, inline filters, and all associated filter holders

• Pump piston seals

• Valve rotors and stators

• All bottles and lids

• All sample vials, microtiter plates, and associated lids and mats

• Columns

• Fuses

• Autosampler needle

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Consumables and Replacement Parts

Solvent System Schematic for the EASY-nLC II Instrument

Solvent System Schematic for the EASY-nLC II Instrument

Table 32

lists the tubing requirements for the EASY-nLC II instrument. The numbers in the

ID column correspond to the numbers in the solvent system schematic shown in

Figure 259 .

7

8

5

6

2

3

4

Kit LC230 includes the Teflon tubing and the check valve assembly required to replace the low-pressure solvent system for one pump. Kit LC243 includes a complete set of flow lines to the mixing Tee, but does not include the mixing Tee itself.

Table 32. Tubing requirements for the EASY-nLC II system

No.

1

Part number

LC212

9

10

11

12

Connections

Pump A outlet to pressure sensor A inlet

Pump S outlet to pressure sensor S inlet

Tubing

PEEK, 300 μm ID, 15 cm length

Pressure sensor A outlet to port 1 of valve A

Pressure sensor S outlet or port 2 of valve S

Pump B outlet to pressure sensor B inlet

Pressure sensor B outlet to port 1 of Valve B stainless steel, prebent stainless steel, prebent

PEEKsil, 50 μm ID, 25 cm length Valve A to flow sensor A

Valve B to flow sensor B

Flow sensor A to mixing Tee

Flow sensor B to mixing Tee

PEEKsil, 20 μm ID, 21 cm length

Fused silica, 10 μm ID, 15 cm length

Mixing Tee to valve S PEEKsil, 30 μm ID, 13 cm length

Autosampler needle connected to port 1 of valve S PEEKsil, 150 μm ID, 55 cm length

ASA model

ASC model

Column Out tubing connected to port 3 of valve S PEEKsil, 30 μm ID, 37 cm length

Waste In line, venting Tee to port 2 of valve W

Port 2 of valve A to check valve A

PEEKsil, 75 μm ID, 50 cm length

Teflon, 500 μm ID, 15 cm length

Port 2 of valve B to check valve B

Tubing from check valve A to solvent bottle A

Tubing from check valve B to solvent bottle B

Teflon, 500 μm ID, 37 cm length

LC215

LC216

LC222

Kit LC243

LC251

LC302

LC260

LC262

Kit,

Low-pressure solvent lines

LC230

13

14

Tubing from check valve A to the waste beaker

Tubing from check valve B to the waste beaker

Tubing from valve W to the waste beaker

Teflon, 500 μm ID, 39 cm length

Teflon, 500 μm ID, 27 cm length LC263

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Consumables and Replacement Parts

Solvent System Schematic for the EASY-nLC II Instrument

Figure 259. Solvent system schematic for the EASY-nLC II system

12

8

13

13

10

1

2

W

6

14

9

1

2

S

6

7

12

Solvent A

Check valves for solvent A

11

1

2

A

6

Solvent B

Check valves for solvent B

11

4

1

2

B

6

Flow sensor A

5

Mixing Tee

4

Flow

↓ sensor

B

6

Waste

1

Pressure sensor

1

1

Pressure sensor

1

3

Pressure sensor

2

PEEK

Stainless steel

PEEKsil

Teflon

Fused silica

Pump S Pump A Pump B

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C

Consumables and Replacement Parts

Solvent System Schematic for the EASY-nLC 1000 Instrument

Solvent System Schematic for the EASY-nLC 1000 Instrument

The pump and valve compartment behind the right panel of the EASY-nLC instrument contains all the flow lines between the components. In addition to pumps and valves, the solvent delivery and waste system includes several filters and four check valves. The two check valves on each solvent line ensure that solvent is drawn from the solvent bottles and ejected to waste without switching valve positions.

Table 33

lists the tubing requirements and identifies the tubing components shown in

Figure 260

. The Low-Pressure Solvent Line Kit (P/N LC230) contains the Tees, check valves, and low-pressure solvent lines to connect one pump (A or B) to the waste system and to its corresponding solvent bottle and six-port rotary valve. The EASY-nLC 1000 Flow Line Kit

(P/N LC570) contains all of these flow lines. The stainless steel tubing is prebent.

Table 33. Tubing requirements for the EASY-nLC 1000 system

2

3

4

No.

1

5

6

7

8

9

D

E

F

Connections

Pump outlet to pressure sensor inlet

Tubing Part number

Stainless steel, 250 μm ID, 150 mm length LC512

Pressure sensor outlet to valve A or B

Pressure sensor outlet to valve S

Stainless steel, 250 μm ID, 150 mm length LC513

Stainless steel, 250 μm ID, 150 mm length LC514 nanoViper, 20 μm ID, 350 mm length LC522 Mixing Tee to valve S, valve A to flow sensor A, valve B to flow sensor B, flow sensor A to mixing Tee

Flow sensor B to mixing Tee

Autosampler needle connected to port 1 of valve S

Column Out tubing connected to port 3 of valve S

Waste In line, venting Tee to port 2 of valve W

Sample loop, 20 μL

Port 2 of valve A to check valve A

Port 2 of valve B to check valve B

Tubing (2) from check valves to solvent bottles nanoViper, 10 μm ID, 180 mm length

PEEKsil™, 150 μm ID, 550 mm length nanoViper, 20 μm ID, 550 mm length nanoViper, 75 μm ID, 550 mm length nanoViper, 250 μm ID, 410 mm length

Teflon™, 500 μm ID, 150 mm length

Teflon, 500 μm ID, 390 mm length

Tubing (2) from check valves to waste beaker Teflon, 500 μm ID, 390 mm length

LC543

LC302

LC560

LC562

LC472 kit LC230 kit LC230 kit LC230

G Tubing from valve W to waste beaker Teflon, 500 μm ID, 330 mm length LC263

316

EASY-nLC Series Troubleshooting and Maintenance Guide Thermo Scientific

C

Consumables and Replacement Parts

Solvent System Schematic for the EASY-nLC 1000 Instrument

Figure 260. Solvent system schematic for the EASY-nLC 1000 system

E E

Waste

7

6

1

2

S

6

9

F

F

8

1

2

W

6

3

Pressure sensor

1

G

Solvent A

Solvent B

4

D

1

2

A

6

2

4

D

1

2

B

6

4

Pressure sensor

1

4

5

2

Pressure sensor

1

Thermo Scientific EASY-nLC Series Troubleshooting and Maintenance Guide

317

C

Consumables and Replacement Parts

Common Replacement Parts

Common Replacement Parts

For proper maintenance of your EASY-nLC instrument, Thermo Fisher Scientific recommends that you maintain an inventory of replacement parts.

Common Replacement Parts for the EASY-nLC II Instrument

Common Replacement Parts for the EASY-nLC 1000 Instrument

Common Replacement Parts for the EASY-nLC II Instrument

Table 34

lists the annual consumption of common replacements parts for the

EASY-nLC II instrument.

Table 34. Common replacement parts for the EASY-nLC II instrument

Description

Pump Piston Seal

Replacement Kit (contains four piston seals and the piston seal tool)

Valve rotor seal for valve serial numbers V-009999 and below (contains one rotor seal)

Valve rotor seal for valve serial numbers V-010000 and above (contains one rotor seal)

Column Out solvent line

Waste In solvent line

Autosampler needle, ASA model

Autosampler needle, ASC model

Valve to flow sensor line

(contains two flow sensor lines)

Replacement filter discs

(contains two 10 μm filter discs)

Part number

LC210

LC224

LC228

LC260

LC262

2

1

LC251 1

LC302 2

LC222 2

LC232

Quantity

4 piston seals (1 per pump) +

1 extra

8 rotor seals (2 per valve)

8 rotor seals (2 per valve)

2 filter discs (1 per solvent bottle)

318

EASY-nLC Series Troubleshooting and Maintenance Guide Thermo Scientific

C

Consumables and Replacement Parts

Common Replacement Parts

Common Replacement Parts for the EASY-nLC 1000 Instrument

Table 35

lists the annual consumption of common replacements parts for the

EASY-nLC 1000 instrument.

Table 35. Common replacement parts for the EASY-nLC 1000 instrument

Description

Pump Piston Seal

Replacement Kit (contains four spring-energized piston seals and the piston seal tool)

Valve rotor seal

(contains one rotor seal)

Column Out solvent line

Waste In solvent line

Autosampler needle, ASC model

Flow sensor filters (contains four flow sensor filters)

Part number

LC510

LC228

LC542

Quantity

4 piston seals

(1 per pump) + 1 extra

8 rotor seals (2 per valve)

LC560 2

LC562 1

LC302 2

4 flow sensor filters

(2 per flow sensor)

Thermo Scientific EASY-nLC Series Troubleshooting and Maintenance Guide

319

D

Quick Reference Guides for Routine Maintenance

This appendix contains a set of quick reference guides for the routine maintenance procedures that you can perform to keep the EASY-nLC system in optimal working condition. These guides are the condensed versions of the procedures in the Routine Maintenance chapter.

If you have replaced a PLF pump in the EASY-nLC II instrument with a PLU pump, follow the instructions in

“Maintaining the PLU Pump Quick Reference Guide” on page 327

to maintain the new PLU pump.

Contents

Using nanoViper Fittings Quick Reference Guide

Maintaining the PLU Pump Quick Reference Guide

Maintaining the PLF Pump Quick Reference Guide

Maintaining the Rotary Valves Quick Reference Guide

Replacing a Damaged Autosampler Needle

Thermo Scientific manual-name

321

Using nanoViper Fittings Quick Reference Guide

For the EASY-nLC 1000 instrument, most of the plumbing connections use nanoViper fittings (see Figure 1

).

Figure 1. nanoViper fitting

PEEK sealing surface

Screw

Removable knurled nut

Connecting nanoViper

Fittings

Even though nanoViper fittings can withstand UHPLC back pressures of up to ~1034 bar (~15 000 psi), they are fingertight fittings, which require only very small torques to seal. To avoid damage by overtightening, follow the instructions in this guide.

Contents

Connecting nanoViper Fittings

nanoViper Tubing Connections

Returning the System to Atmospheric Pressure

To connect a nanoViper fitting to a receiving port

1. Insert the nanoViper fitting into the receiving port. Then turn the screw clockwise until you feel resistance.

2. Using the black knurled nut, turn the screw clockwise to an angle between 0 and 45 degrees (1/8-turn).

3. Run the system leak test (see “Running a System Leak Test” on page 218 ).

When the leak test ends, the system is at atmospheric pressure.

IMPORTANT To extend the lifetime of the nanoViper fittings, open and close connections at atmospheric system pressures only. Opening and closing connections at high system pressures can reduce the lifetime of the fitting system.

4. If the Leaks script fails because the new connection is not leak tight, use the black knurled nut to turn the screw up to an additional 45 degrees. Do not turn the screw beyond an angle of 90 degrees from where you felt the initial resistance.

IMPORTANT To prevent damage to the sealing surface of the nanoViper fitting, take care not to overtighten the fingertight nanoViper fitting.

© 2012 Thermo Fisher Scientific Inc.

All rights reserved.

nanoViper

Tubing

Connections

Figure 2 shows the nanoViper tubing connections in the EASY-nLC 1000 instrument.

• Port 6 of valve A to flow sensor A inlet

• Port 6 of valve B to flow sensor B inlet

• Flow sensor A outlet to mixing Tee inlet

• Flow sensor B outlet to mixing Tee inlet

• Mixing Tee outlet to port 4 of valve S

• Sample loop connected to ports 2 and 5 of valve S

• Column Out line connected to port 3 of valve S and the HPLC union

• Waste In line connected to port 2 of valve W and the HPLC union or the venting Tee

Figure 2. nanoViper tubing connections in the EASY-nLC 1000 instrument

Valve A

2 6

Sample loop

2

Valve S

1

6

Flow sensor A

Mixing Tee

3

4

5

Column Out line

2

Valve B

6

Waste In line

2

Valve W

6

Flow sensor B

324

Returning the System to

Atmospheric

Pressure

Upstream of

Valve S

Solvent Lines

Before you disconnect a nanoViper fitting from its receiving port, always return the solvent line to atmospheric pressure. The high-pressure solvent lines in the EASY-nLC solvent system can be divided into two categories based on their position relative to valve S.

Upstream of Valve S Solvent Lines

Downstream of Valve S Solvent Lines

The following solvent lines (numbered from left to right in

Figure 3 ) are in the flow path upstream of valve S.

#

1

2

3

Solvent line

Port 6 of valve A to flow sensor A inlet

Port 6 of valve B to flow sensor B inlet

Flow sensor A outlet to mixing Tee inlet

#

4

5

6

Solvent line

Flow sensor B outlet to mixing Tee inlet

Mixing Tee outlet to port 4 of valve S

Sample loop connected to ports 2 and 5 of valve S

To return the solvent lines that are upstream of valve S to atmospheric pressure

1. On the touch screen, press Home > Overview.

2. Using the valve controls, set the valves to the following positions.

Valve

A and B

Position

Center

Effect

Maintains the pressure between the pump A and valve A during the

Analytical Column Equilibration script.

Vents the system pressure downstream of valve A.

S and W 1–6

Figure 3 shows the flow path when valves A and B are Centered and valves S and W are in position 1–6.

Figure 3. Solvent lines that are downstream of valves A and B and upstream of valve S, numbered 1–6

1

No flow

2

3

1

A

4

6

5

Center position

2

No flow

2

3

1

B

4

6

5

Center position

3

Mixing Tee

4

2

3

1

S

4

6

5

5

6

2

3

1

W

6

5

4

Waste beaker

325

Downstream of

Valve S

Solvent Lines

The following solvent line connections are in the flow path downstream of valve S:

• Column Out tubing connected to port 3 of valve S and the HPLC union (P/N SC900)

• Waste In tubing connected to port 2 of valve W and the HPLC union or the venting Tee

To return the solvent lines that are downstream of valve S to atmospheric pressure

1. On the touch screen, press Home > Overview.

2. Using the valve controls, set the valves to the following positions.

Valve

S

W

Position

Center

1–6

Effect

Maintains the system pressure upstream of valve S.

Vents the system pressure downstream of valve S.

Figure 4 shows the Column Out and Waste In solvent lines that are downstream of valve S.

Figure 4. Solvent lines that are downstream of valve S

2

3

1

S

4

6

5

Column Out line

Waste In line

2

3

1

W

6

5

4

Waste beaker

326

Maintaining the PLU Pump Quick Reference Guide

Replace the piston seal when you detect a leak in the pump.

Contents

Removing a Used Piston Seal

Installing a New Piston Seal

Replacing a piston seal requires these tools and materials.

Tools

3 mm hex wrench

Piston seal tool

1/4 in. open-ended wrench

#2 Phillips head screwdriver

Materials

Pipette

Powder-free safety gloves

LC/MS-grade methanol

LC/MS-grade acetonitrile

Piston seal, P/N LC510

Removing a Used Piston Seal

1. Set the pump piston to the fully retracted position as follows: a. On the touch screen, press Maintenance > Scripts.

b. In the Category list, select Prepare.

c. In the Name list, select Purge Solvent.

d. Press the Parameters tab. e. Press Purge Iterations and enter 0.

Figure 1. Purge Solvent script set to 0 iterations

2. Exit the EASY-nLC 1000 system and turn off the power.

3. Using a #2 Phillips head screwdriver, with a quarter-turn loosen the three captive screws that secure the right side panel to the instrument. Then remove the panel.

CAUTION Wear powder-free gloves when handling the wetted components of the LC system

.

4. Remove the used piston seal as follows: a. Using a 1/4 in. open-ended wrench, remove the stainless steel tubing connected to the pump head.

Figure 2. Stainless steel fitting removed from the pump head

Stainless steel tubing b. Using a 3 mm hex wrench, remove the two screws that secure the pump head to the pump body.

Figure 3. Pump head removed from the pump body f. Select the check box for the appropriate pump.

g. Press Start.

The piston moves downward to the 140 μL position.

c. Clean the visible portion of the piston with a lint-free tissue soaked in LC/MS-grade methanol and visually inspect the piston for scratches. Ensure that no solvent runs into the pump.

d. Remove the old seal from the pump head with great care. Do not use any metal tools, as they can scratch the inside of the pump head and generate leaks.

Installing a New Piston Seal

1. With the spring side of the piston seal spring facing away from the piston seal tool, mount the new piston seal onto the piston seal tool.

2. Using a pipette, remove the air from the piston seal spring by carefully filling the rim of the piston seal with methanol.

Figure 4. Filling the piston seal with methanol d. Pipette the appropriate solvent into the pump head.

Figure 7. Adding solvent to the pump head

3. Place the pump house on the guide rods and evenly press it all the way down to insert the seal and then gently remove the house when the seal is in position.

Figure 5. Inserting the piston seal into the pump head

4. Reassemble the pump head on the pump using the two screws and a 3 mm hex wrench. Do not connect the stainless steel tubing to the pump head.

5. Turn on the EASY-nLC system and log in as an administrator.

6. Fill the pump head with the appropriate solvent as follows: a. Press Home > Overview.

b. Press the pump icon for the pump that you want to control.

The Position readback displays the current pump position from 0 μL (empty) to 140 μL (full). c. Set the flow rate to 300 μL/min and the volume to 140 μL.

Then press Start.

The piston moves up to the top of the pump head.

Figure 6. Pump dialog box e. Set flow rate to –300 μL/min and the volume to 140 μL.

Then press Start.

The piston moves down, drawing solvent into the pump head. Make sure the pump head is filled with solvent during the entire retraction of the piston.

CAUTION Be careful not to spill solvent onto the printed circuit board behind the LED panel.

7. Using a 1/4 in. open-ended wrench, reconnect the tubing to the

pump head (see Figure 2 on page 327

).

8. Reinstall the right side panel.

9. Reset the usage counter for the pump as follows: a. Press Maintenance > Devices. b. Select the pump from the list of devices.

c. Press the Summary tab.

d. Press Reset.

The value in the Intermediate Volume box resets to 0.

10. To draw fresh solvent through the solvent lines and to remove air, do the following: a. Open the Parameters page for the Purge Solvent script as described in

step 1 on page 327 .

b. In the Purge Iterations box, enter 10.

c. Select the check box for the appropriate pump. d. Press Start and wait for the system to perform 10 purge cycles.

11. To remove air from the pump head, do the following: a. In the Name category on the Maintenance > Scripts page, select Flush Air.

b. Press the Parameters tab.

c. In the Flush Threshold [μL] box, enter 12.

d. Press Start.

e. Wait for the script to finish.

12. Run the Leaks script for the pump with the new piston seal a. On the Maintenance > Scripts page, select Test in the

Category list, and then select Leaks in the Name list.

b. Press the Parameters tab.

c. Select the check box for the pump with the new seal.

d. Press Start.

13. . When the instrument is leak tight, it is ready for use.

328

Maintaining the PLF Pump Quick Reference Guide

Replace the piston seal when you detect a leak in the pump.

Contents

1.

Removing the Pump Head and Cleaning the Piston

2.

Removing the Worn Piston Seal

3.

Installing a New Piston Seal

4.

Priming the Pump

5.

Preparing the Instrument for Operation

CAUTION Wear powder-free safety gloves when working with solvents and handling the wetted components of the instrument.

Replacing a piston seal requires these tools and materials.

Tools Materials

3 mm hex wrench

Piston seal tool

Pipette

Powder-free safety gloves

1/4 in. open-ended wrench LC/MS-grade methanol

#2 Phillips head screwdriver LC/MS-grade acetonitrile

Piston seal, P/N LC210 (includes the piston seal tool)

Removing the Pump Head and Cleaning the Piston

1. Set the pump piston to the fully retracted position as follows: a. On the Maintenance > Scripts page, Select Prepare in the

Category list and then select Purge Solvent in the Name list.

Press the Parameters tab. In the Purge Iterations box, enter 0.

Select the check box for the appropriate pump.

b. Press Start. The piston moves down to the 140 μL position.

2. Exit the EASY-nLC II system and turn off the power.

3. Using a #2 Phillips head screwdriver, with a quarter-turn loosen the three captive screws that secure the right side panel to the instrument. Then remove the panel.

4. Remove the tubing connected to the pump head as follows:

• If a PEEK fitting is connected to the pump head, use a

13 mm open-ended wrench, to remove it.

Figure 1. PEEK fitting on the externally threaded pump head

• If a stainless steel fitting is connected to the pump head, use a

1/4 in. open-ended wrench to remove it.

Figure 2. Stainless steel fitting on the internally threaded pump head

Stainless steel fitting connected to an internally threaded pump head

5. Using a 13 mm open-ended wrench, remove the pump head.

Figure 3. Removing the pump head

Pump head

6. Place the piston in the fully extended position as follows: a. Turn on the instrument and log in as an administrator.

b. Press Home > Overview, and then press the pump icon for the pump you want to control. The Pump dialog box opens.

c. Set the flow rate to 300 μL/min and the volume to 140 μL.

d. Press Start.

7. Clean the piston with the lint-free tissue soaked in methanol, and visually inspect the piston for any scratches.

CAUTION Take care to avoid solvent drips onto the pump PCB.

8. Return the piston to the fully retracted position as follows: a. In the Pump dialog box, set the flow rate to – 300 μL/min and the dispense volume to 140 μL. b. Press Start.

9. When the piston is fully retracted, close down the EASY-nLC system and turn off the power.

PEEK fitting connected to an externally threaded pump head

Removing the Worn Piston Seal

1. Clean the piston seal tool in a beaker filled with 100% methanol.

Figure 4. Two-component piston seal tool for the PLF model pump

2. Insert the piston seal tool into the worn piston seal and pull the piston seal out of the pump head.

Figure 5. Using the piston seal tool to remove the worn piston seal

Installing a New Piston Seal

1. Insert the guide tube into the pump head flange.

Figure 6. Guide tool inserted into the pump head flange

Guide tool

Worn piston seal

2. Using a pipette, fill the groove in the piston seal with methanol.

3. Place the new seal with the groove facing up on the piston seal tool.

Figure 7. Piston seal mounted onto the piston seal tool

4. Insert the piston seal tool into the guide tool.

Figure 8. Inserting the piston seal tool into the guide

5. Push the piston seal tool into the guide until you feel resistance.

6. Remove the guide tube and the piston seal tool, and then check that the piston seal is properly seated.

7. Using a 13 mm open-ended wrench, reconnect the pump head to the pump body. Do not connect the stainless steel tubing to the pump head.

330

Priming the Pump

1. Turn on the EASY-nLC instrument and log in as an administrator.

2. Fill the pump head with the appropriate solvent as follows: a. Press Home > Overview.

b. Press the pump icon for the pump that you want to control.

c. Set the flow rate to 300 μL/min and the volume to 140 μL.

Then press Start.

The piston moves upward to the top of the pump head.

d. Pipette the appropriate solvent into the pump head.

e. Set the flow rate to –300 μL/min and the volume to 140 μL.

Then press Start.

The piston moves down, drawing solvent into the pump head. Make sure the pump head is filled with solvent during the entire retraction of the piston.

CAUTION Be careful not to spill solvent onto the printed circuit board behind the LED panel.

3. Using a 1/4 in. open-ended wrench, reconnect the tubing to the pump head.

Preparing the Instrument for Operation

1. Reinstall the right panel.

2. Reset the usage counter for the pump as follows: a. Press Maintenance > Devices. b. Select the pump from the list of devices.

c. Press the Summary tab.

d. Press Reset.

The value in the Intermediate Volume box resets to 0.

3. To draw fresh solvent through the solvent lines and to remove air, do the following: a. Open the Parameter page for the Purge Solvent script.

b. In the Purge Iterations box, enter 10.

c. Select the check box for the appropriate pump. d. Press Start and wait for the system to perform 10 purge cycles.

4. To flush air from the pump head, do the following: a. In the Name category on the Maintenance > Scripts page, select Flush Air.

b. Press the Parameters tab.

c. In the Flush Threshold [μL] box, enter 10.

d. Press Start.

e. Wait for the script to finish.

5. Run the Leaks script for the pump as follows: a. On the Maintenance > Scripts page, select Test in the

Category list, and then select Leaks in the Name list.

b. Press the Parameters tab.

c. Select the check box for the pump with the new seal.

d. Press Start.

6. When the instrument is leak tight, it is ready for use.

Maintaining the Rotary Valves Quick Reference Guide

T

Replace the rotor seal when you detect a leak or blockage in the valve.

Contents

Replacing a Rotor Seal

Replacing the Stator

CAUTION Wear powder-free safety gloves when working with solvents and handling the wetted components of the instrument.

Replacing a piston seal requires these tools and materials.

Tools Materials

• 9/64 in. hex wrench • Rotor seal, P/N LC228

• 1/4 in. open-ended

(EASY-nLC system valves with wrench serial numbers V-010000 and above)

• #2 Phillips head screwdriver

• Rotor seal, P/N LC224

(EASY-nLC II system valves with serial numbers below V-009999)

• LC/MS-grade methanol

• Powder-free safety gloves

Replacing a Rotor Seal

1. Close down the EASY-nLC system, and then turn off the power to the instrument.

2. Using a #2 Phillips screwdriver, with a quarter-turn loosen the three captive screws that secure the panel to the right side of the

EASY-nLC instrument, and then remove the panel.

3. Using a 9/64 in. L-hex wrench, remove the two hex screws that secure the stator to the valve assembly, and then pull the stator away from the valve driver.

4. Remove the rotor seal from the valve driver.

Figure 1. Rotor seal mounted on the valve driver

5. Clean the stator with a lint-free tissue or Q-tip soaked in methanol.

Figure 2. Cleaning the stator

6. Carefully mount the new rotor onto the valve driver, ensuring that the rotor sealing surface (engraved flow passage), is facing out.

7. Mount the stator onto the valve driver.

8. Using a 9/64 in. L-hex wrench, tighten the two hex screws a little at a time by shifting from one screw to the other and back again until the screws are evenly torqued.

9. Reconnect the right side panel.

10. Turn on the EASY-nLC instrument and log in as an administrator.

11. Reset the usage counter for the valve as follows: a. Press Maintenance > Devices. b. Select the valve from the list of devices.

c. Press the Summary tab.

d. Press Reset.

The value in the Rotor Shifts box resets to 0.

12. If you replaced the rotor seal in Valve A or B, flush air from the system as follows: a. On the Maintenance > Scripts page, select Prepare in the

Category list. Then select Flush Air in the Name list. b. Select the pump that is connected to the affected valve.

c. Set the flush volume threshold to 10 μL for the EASY-nLC II or 12 μL for the EASY-nLC 1000.

d. Press Start.

e. Wait for the script to finish.

13. Run the Leaks script as follows: a. Select Test in the Category list. Then select Leaks in the

Name list.

b. Do one of the following:

• If you replaced the rotor seal in Valve A or B, run the

Leaks script for the affected valve.

• If you replaced the rotor seal in Valve S or W, run the

Leaks script for the system.

When the system is leak tight, the instrument is ready for use.

Replacing the Stator

1. Close down the EASY-nLC system, and then turn off the power to the instrument (see

“Closing Down the EASY-nLC

Instrument” on page 21 ).

2. Using a #2 Phillips screwdriver, loosen the three quarter-turn screws that secure the panel to the right side the EASY-nLC instrument, and then remove the panel.

3. Disconnect the solvent lines from the valve as follows:

• Use a 1/4 in. open-ended wrench to remove stainless steel fittings.

• Use the black nanoViper knurled nut to remove nanoViper fittings.

4. Using a 9/64 in. L-hex wrench, remove the two hex screws that secure the stator to the valve driver, and then pull the stator away from the valve driver.

5. Mount the new stator onto the valve driver.

6. Using a 9/64 in. L-hex wrench, tighten the two hex screws a little at a time by shifting from one screw to the other and back again until the screws are evenly torqued.

7. Reconnect the solvent lines to the valve as follows:

• Use a 1/4 in. open-ended wrench to tighten stainless steel fittings.

• Use the black nanoViper knurled nut to tighten nanoViper fittings.

8. Reconnect the right side panel to the instrument. Tighten the three screws with a #2 Phillips screwdriver.

9. Turn on the EASY-nLC instrument and log in as an administrator.

10. If you replaced the stator in Valve A or B, run the Flush Air script for the valve with the replacement stator.

11. If you replaced the stator on Valve A or B, run the Leaks script for the valve with the replacement stator. If you replaced the stator on

Valve S or W, run the Leaks script for the system.

332

Replacing a Damaged Autosampler Needle

T

Replace the autosampler needle when it is bent or clogged.

Contents

Removing the Damaged Autosampler Needle

Installing a New Autosampler Needle

CAUTION Wear powder-free safety gloves when working with solvents and handling the wetted components of the instrument.

Replacing a damaged needle requires these tools and materials.

Tools Materials

• 1/4 in. open-ended wrench • Powder-free safety gloves

• #2 Phillips head screwdriver • ASA autosampler needle,

P/N LC251

• ASC autosampler needle,

P/N LC302

Removing the Damaged Autosampler Needle

To remove the autosampler needle

1. Using a #2 Phillips screwdriver, with a quarter-turn loosen the three captive screws that secure the panel to the right side the

EASY-nLC instrument, and then remove the panel.

2. Using a 1/4 in. open-ended wrench, unscrew the nut that secures the autosampler needle to port 1 of valve S. Then remove the fitting from the port.

Figure 1. Valve S solvent line connections for the EASY-nLC II system

Autosampler needle

Column Out line

2

3

3

2

Valve S

1

1

6

5

4

4

From the mixing Tee

6

5

From the pressure sensor

Figure 2. Valve S solvent line connections for the EASY-nLC 1000 system

1

Autosampler needle

2

6

From the pressure sensor

5

3

Column Out line

From the mixing Tee

4

3. Remove the nut, ferrule, and sleeve from the needle tubing.

Figure 3. Autosampler tubing with a nut, ferrule, and sleeve at one end

4. Move the z-axis needle holder to an accessible location within the tray compartment by doing one of the following:

• For the ASA model, go to

step 5 .

• For the ASC model, go to

step 6 .

5. For the ASA model, move the z-axis needle holder to position A1 as follows: a. Press Home > Overview.

b. Press the autosampler icon.

c. The Autosampler dialog box opens.

d. In the XYZ Robot area, select position A1 in the Well box.

Figure 4. Autosampler direct control dialog box e. Press Goto.

f. Go to step 7

.

6. For the ASC model, move the z-axis needle holder to position W1 as follows: a. Press Home > Overview. b. Press the autosampler icon.

The Autosampler dialog box opens. c. In the XYZ Robot area, press Go to Calibration.

The Tools page of the Autosampler view on the

Maintenance > Devices page opens.

Figure 5. Tools page of the Autosampler view on the Maintenance >

Devices page

Figure 7. Autosampler needle connected to the z-axis needle holder

ASA z-axis needle holder ASC z-axis needle holder d. In the Manipulator area, press Reset.

The z-axis needle holder moves to the W1 position.

7. Using a #2 Phillips screwdriver, loosen the three quarter-turn screws that secure the panel to the left side of the EASY-nLC instrument, and then remove the panel.

8. Do one of the following:

• For the ASC model, remove the black needle holder from the slot in the panel that separates the solvent system hardware from the autosampler compartment. Removing the fitting requires some gentle movement from both sides of the plate.

Figure 6. Removing the black needle holder from the slot in the panel

Black needle holder

Slot in the metal plate

Installing a New Autosampler Needle

To install an new autosampler needle

1. Mount the new autosampler needle in the z-axis holder. Check that the small spring is situated between the PEEK nut and the plastic stop.

2. Do one of the following:

• For the ASC autosampler, go to step

step 3 .

• For the ASA autosampler, go to step

step 4 .

3. For the ASC autosampler, do the following: a. Guide the end of the needle that connects to Valve S through the large hole in the metal plate that separates the tray compartment from the solvent system compartment. b. Install the black needle holder in the side plate.

4. For the ASA autosampler, slide the valve end of the tubing through the small plastic holder on the back of the touch-screen monitor.

Figure 8. Holder on the back of the touch-screen monitor

• For the ASA model, remove the white plastic needle guide on the back of the touch-screen monitor.

9. Loosen the nut that is connected to the z-axis needle holder by turning it counterclockwise. Then carefully pull the autosampler needle upward and away from the holder.

Holder

5. Connect the needle to port 1 of Valve S as follows: a. Slide the provided blue sleeve and metal fittings onto the tubing. b. To ensure a zero dead volume connection, hold the blue sleeve and the tubing firmly against the bottom of the valve port, and then tighten the fitting with a 1/4 in. open-ended wrench.

6. Recalibrate the needle position.

7. Press Maintenance > Scripts.

8. In the Category list, select Prepare.

9. In the Name list, select Purge Solvent. Then run 2 iterations of the Purge Solvent script for Pump S.

10. In the Name list, select Flush Air. Then run the Flush Air script for Pump S with a flush volume threshold of 10 μL for the

EASY-nLC II system or 12 μL for the EASY-nLC 1000 system.

334

I

Index

A

Acquisition Server errors

189

adapter plate, replacing

ASA autosampler

272

ASC autosampler

271

air, removing from pump heads

33

Analytical Column Equilibration script

36

application version

20

ASA autosampler adapter plate, replacing

272

description

4

needle, description of

14

See also

autosampler

upgrading to ASC autosampler (service repair)

128

ASA cooler, replacing (service repair)

140

ASC autosampler adapter plate, replacing

271

description

3

needle, description of

14

replacing (service repair)

122

See also

autosampler

ASC cooler, replacing (service repair)

137

autosampler calibrating XYZ robot

269 –

286

checking sample pickup, schedule

28

description

2

direct control

210

needle blocked

262

description

14

replacing

84 –

88

Sample Pickup script

263

Torque script

49

tray compartment, accessing

2

troubleshooting tips

171

B

back panel, instrument

16

back pressure testing

48

troubleshooting maximum pressure spikes during gradient

180

system blockage

249

too high during sample loading and column equilibration

178

Back Pressure script

48

backup ring

8

beaker, waste

2

blank injections, troubleshooting

183

– 184

, 263

blind fitting, EASY-nLC 1000 instrument

218

blockage, troubleshooting

249

C

cables, internal connected to EASY-nLC 1000 pump

152

cooler bus cable

123

LVDS cable

108 ,

114 ,

116

monitor cable

114

P-Bus communication cables

118 ,

158

PC box cable connections

108

pressure sensor communication cable

119

USB cable

114

calibrating autosampler XYZ robot

269

286

flow sensors

51

plates

280

pressure sensors

52

carryover specification

23

troubleshooting tips

194

check valves description

11

replacing

80

Thermo Scientific EASY-nLC Series Troubleshooting and Maintenance Guide 335

Index: D checking check valves for leaks

29 ,

248

sample pickup

263

solvent levels

27

system for leaks

44

clogged column or system, troubleshooting

178

closing down the EASY-nLC instrument

21

Column Out line checking for blockage

253

clogged

178

, 180

tubing ID

249

column, clogged

178

compliance

FCC

iii

regulatory

iii

WEEE

ix

compliance, electromagnetic

v

, vii

computer, integrated location

15

out of memory

202

PC box, replacing (service repair)

108

consumables

318

contact closure

MS Connection script

39

troubleshooting

172

contamination declaration

301

preventing

205

cooler PCB

132

cooler temperature checking

28

direct control

211

D

daily maintenance

27

Declaration of Conformity

v ,

vii

Declaration of Contamination

301

devices managing the Devices list

105

usage counters

56

Direct Infusion script

53

E

EASY-nLC 1000 instrument (service repairs)

ASC autosampler, replacing

122

127

ASC cooler, replacing

137 –

139

high-pressure valves, replacing

147

– 149

monitor, replacing

113

– 116

PC box, replacing

108

– 112

PLU pump, replacing

150 –

153

pump PCB, replacing

163 –

166

336

EASY-nLC Series Troubleshooting and Maintenance Guide

EASY-nLC II instrument (service repairs)

ASA cooler, replacing

140 –

146

ASC autosampler, replacing

122

127

ASC cooler, replacing

137 –

139

autosampler, upgrading ASA to ASC

128 –

136

monitor, replacing

113

– 116

PC box, replacing

108

– 112

PLF pump, replacing

154

pressure sensor, replacing

117 –

121

pump PCB, replacing

167 –

168

electromagnetic compatibility

iii

Exit button

21

F

failure messages

216

fan cables

145

Fast Inspection option

52

FCC compliance

iii

field service repairs

See

EASY-nLC II instrument or EASY-nLC 1000 instrument (service repairs)

filling a pump with solvent

206

flow rate determining appropriate analytical column

36

precolumn

34

direct control

205

measuring

10

flow sensor cable

152 ,

163 ,

166

flow sensors calibration script

51

checking for blockage

255

, 259

description

10

maintenance schedule

29

replacing

EASY-nLC 1000 instrument

92

EASY-nLC II instrument

96

Flush Air script

33

fuse, replacing

58

G

grounding requirements

268

H

hard drive, replacing

100

HPLC union

218

Thermo Scientific

I

inline filters, replacing in the EASY-nLC 1000 instrument

81

Inspection Only check box

52

instrument startup, troubleshooting

172

interface layout

19

IP address, support server

289

Isocratic Flow script

38

L

L brackets, pressure sensor

91

,

120

,

157

Leaks script

44

46

leaks, checking for

28

,

44

– 46 ,

218

– 247

log book

55

Logo icon, Thermo Scientific

20

loop, sample, replacing

89

lower case letters, cause of Acquisition Server error

189

low-pressure solvent filters

29

LVDS cable, internal

114 ,

116

M

maintenance daily

27

quarterly

28

scripts

31

weekly

28

yearly

29

manuals, EASY-nLC

xx

mass spectrometer, contact closure

MS Connection script

39

troubleshooting

172

, 266

memory stick, correct format

202

message.log file

292

messages, failure

216

mixing Tee checking for blockage

254

installing in the EASY-nLC II instrument

135

monitor cable, internal

114

monitor, replacing (service repair)

113

motor cable

165 –

168

MS Connection script

39

N

nanoViper fittings

EASY-nLC 1000 solvent lines

316

using

83 ,

323

needle arm

135

needle guide, white plastic

115

– 116

needle position, calibrating

278 –

286

needle, autosampler

14

network access, troubleshooting

264

Thermo Scientific

P

parts, common replacement

318

password, initial

20

P-Bus communication cables connecting to cooler PCB

132 ,

139

connecting to pump

166

connecting to pump PCB

121

, 168

connecting to XYZ-axis PCB

132

disconnecting from cooler PCB

129 ,

137

disconnecting from pump

152 ,

163

disconnecting from pump PCB

118

, 158

, 167

disconnecting from XYZ-axis PCB

129

inserting P-Bus connector

131

PCBs connected to

132

P-Bus connector

131

PC box cables connected to

108

replacing (service repair)

108

Terminated label

131

PCB shield

164 ,

166

performance specifications

23

pink autosampler icon

201

pump icons

199

valve icon

200

piston seals function

8

replacing in the PLF model pump

62

in the PLU model pump

67

plate formats creating

275

deleting

277

selecting

274

plates, calibrating

280

plug, EASY-nLC II instrument

218

port 22, firewall setup for

287

– 288

Power Down button

22

power, turning off

22

preparing autosampler for the calibration routines

278

system for daily use

27

pressure sensor cable

152 ,

163 ,

166

pressure sensors communication cable

119 ,

121 ,

167

description

10

grounding cable connecting to PCB (service repair)

118 ,

121 ,

168

replacing

EASY-nLC 1000 instrument

(routine maintenance)

90

EASY-nLC II instrument (service repair)

117

Index: I

EASY-nLC Series Troubleshooting and Maintenance Guide

337

Index: Q pump direct control

205

manual control

71

, 205

PLF, EASY-nLC II instrument description

8

maintaining

62

replacing (service repair)

154

PLU, EASY-nLC 1000 instrument description

8

maintaining

67

replacing (service repair)

150

volume pumped usage counter

56

Pump dialog box

206

Pump Mounting Kit

154 ,

160

pump PCB, replacing (service repair)

EASY-nLC 1000 instrument

163

– 166

EASY-nLC II instrument

167 –

168

Purge Solvent script

32

Q

quick reference guides autosampler needle, replacing

333

EASY-nLC 1000 pump, maintaining

327

EASY-nLC II pump, maintaining

329

nanoViper fittings, using

323

valves, maintaining

331

R

regulatory compliance

iii

remote support

287

replacement parts, common

318

Reset Pressure Sensor script

52

rotary valves

See

valves

rotor seal, replacing

77

S

safety calibrating the autosampler

278

precautions for the EASY-nLC instrument

xxi

standards (EMC)

vii

safety standards

iii

safety standards (EMC)

v

sample pickup check

263

Pickup script

42

sample loop, replacing

89

schedule, maintenance

27

338

EASY-nLC Series Troubleshooting and Maintenance Guide scripts

Analytical Col Equilibration

36

Autosampler Torque

49

Back Pressure

48

Flow Sensors

51

Flush Air

33

Isocratic Flow

38

Leaks

44

Maintenance

31

MS Connection

39

Purge Solvent

32

Reset Pressure Sensor

52

Sample Pickup

42 ,

263

Valve Check

47

Valve Tune

50

seals piston description

8

replacing in the PLF model pump

62

in the PLU model pump

67

Secure Shell (SSH) network protocol

287

serial numbers, device

106

service log

55

service repairs

See

EASY-nLC II instrument or EASY-nLC 1000 instrument (service repairs)

sharp objects, autosampler needle

278

sluggish system operation

202

software, upgrading the touch-screen application

128

,

296

solvent system components behind the right side panel

5

schematic

EASY-nLC 1000 system

317

EASY-nLC II system

315

solvents checking levels

27

viscosity table

311

specifications performance

23

technical

24

spray, unstable

196

SSH Connection Error message box

292

stator cleaning

75

replacing

79

support collar

148

support server, IP address

289

swept volume, extra

176

system overload

202

Thermo Scientific

T

technical specifications

24

technical support, contacting

xxiii

Telnet window

266

temperature direct control

211

maintaining stable

284

Terminated label, PC box

131

touch-screen software description

18

upgrading

128

, 296

transport instructions

299

tray compartment, accessing

2

troubleshooting autosampler

171

contact closure

172

identifying and locating leaks

218

instrument startup

172

network access

264

system blockage

249

turning off the EASY-nLC instrument

21

U

union, HPLC

218

upgrading the touch-screen software

128

,

296

URLs consumable parts

xxiii

customer manuals

xx

usage counters, device

56

USB cable, internal

114

USB ports, EASY-nLC instrument

17 ,

293

USB storage device, copying system files

293

user interface layout

19

V

Valve Check script

47

valve position description

12

manual control

204

Valve Tune script

50

valves check valves description

11

replacing

80

checking for blockage valve B

257

valves A, S, and W

251

checking for leaks valve B

223

rotary description

12

maintaining

74

replacing

147

versions, application

20 ,

297

vials extra vial positions, calibrating

285

plate format, calibrating

280

viscosities, solvent

311

W

wash bottles calibrating W4 bottle with insert

285

description

2

waste beaker, description

2

Waste In line checking for blockage

253

weak signal from detector

183

WEEE compliance

ix

weight, EASY-nLC instrument

25

well plates calibrating

280

plate formats, managing

274

X

Xcalibur data system, errors reported by

189

XYZ robot calibrating

278

286

direct control

210

XYZ-axis PCB

123 ,

127 ,

129 ,

132

XYZ-axis PCB bus cable

123

Index: T

Thermo Scientific EASY-nLC Series Troubleshooting and Maintenance Guide

339

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